Inhibitors of egfr and methods of use thereof

ABSTRACT

The application relates to compounds, or pharmaceutically acceptable salts, hydrates, solvates, or stereoisomers thereof, which modulate the activity of EGFR, a pharmaceutical composition comprising said compounds, and methods of treating or preventing a disease in which EGFR plays a role.

RELATED APPLICATION

This application claims priority to U.S. Provisional Application No.62/688,057 filed Jun. 21, 2018, the contents of which is herebyincorporated by reference in its entirety.

GOVERNMENT SUPPORT

This invention was made with government support under grant number R01CA201049 awarded by the National Institutes of Health. The governmenthas certain rights in the invention.

BACKGROUND

The epidermal growth factor receptor (EGFR, Erb-B1) belongs to a familyof proteins involved in cell proliferation. EGFR overexpression ispresent in at least 70% of human cancers, such as non-small cell lungcarcinoma (NSCLC), breast cancer, glioma, and prostate cancer. TheEGFR-TK is therefore widely recognized as a target for the design anddevelopment of therapies that can specifically bind and inhibit tyrosinekinase activity and its signal transduction pathway in cancer cells, andthus can serve as diagnostic or therapeutic agents.

EGER tyrosine kinase inhibitors (TKIs) are effective clinical therapiesfor EGFR mutant advanced non-small cell lung cancer (NSCLC) patients.However, the vast majority of patients develop disease progressionfollowing successful treatment with an EGFR TKI. The most commonmechanism of acquired resistance is a secondary mutation T790M, whichleads to an increase in ATP affinity, thus making it more difficult forreversible EGFR TKIs gefitinib and erlotinib to bind the EGFR domain.Covalent EGFR inhibitors have emerged as strategies to inhibit EGFRT790M containing cancers. Afatinib is a potent inhibitor of both mutantand wild type (WT) EGFR, but is only effective in EGFR TKI naive EGERmutant cancers, has a RR of <10% in patients with NSCLC resistant togefitinib or erlotinib, and suffers from toxicities from inhibition ofWT EGFR. Other irreversible EGFR inhibitors, such as WZ4002, CO-1686,and AZD9291, overcome many of the limitations of afatinib. They are notonly more potent on EGFR T790M, but also selectively inhibit mutant overWI EGFR.

However, all current EGFR TKIs target the ATP binding site, and arerendered impotent by the C797S mutation arising in treated patients.Cetuximab, an anti-EGFR antibody that blocks receptor dimerization isnot effective in EGER-mutant NSCLC, because mutational activation of thekinase is effectively “downstream” of receptor dimerization. Hence,alternative strategies to inhibit EGFR are needed. Thus, there is a needfor novel and potent small molecule EGFR inhibitors with alternativemechanisms of action targeting mutant EGFR. The present applicationaddresses the need.

SUMMARY

The present application relates to benzodiazepine allosteric inhibitorsof EGFR that are capable of inhibiting drug resistant forms of EGFR. Theapplication features methods of treating or preventing a disease inwhich EGFR plays a role in a subject in need thereof by administering tothe subject a therapeutically effective amount of a compound describedherein, or a pharmaceutically acceptable salt, hydrate, solvate, orstereoisomer thereof. The methods of the application can be used totreat or prevent diseases in which EGFR plays a role by inhibiting thekinase activity of EGFR.

A first aspect of the application relates to a compound of Formula I:

or a pharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof, wherein each of the variables in Formula I is described hereinin detail below.

Another aspect of the present application relates to a pharmaceuticalcomposition comprising a compound of Formula I, or a pharmaceuticallyacceptable salt, hydrate, solvate, or stereoisomer thereof, and apharmaceutically acceptable carrier. In another aspect, thepharmaceutical composition further comprises a second agent thatprevents EGFR dimer formation, and a pharmaceutically acceptablecarrier.

Another aspect of the present application relates to a method ofinhibiting a kinase (e.g., EGFR). The method comprises administering toa subject in need thereof an effective amount of a compound of FormulaI, or a pharmaceutically acceptable salt, hydrate, solvate, orstereoisomer thereof. In one aspect, the method further comprisesadministering to the subject a second agent that prevents EGFR dimerformation.

Another aspect of the present application relates to a method oftreating or preventing a disease (e.g., a disease in which EGFR plays arole). The method comprises administering to a subject in need thereofan effective amount of a compound of Formula I, or a pharmaceuticallyacceptable salt, hydrate, solvate, or stereoisomer thereof. In anotheraspect, the method further comprises administering to the subject asecond agent that prevents EGFR dimer formation.

Another aspect of the present application relates to a method oftreating or preventing a disease resistant to an EGFR targeted therapy,such as a therapy with gefitinib, erlotinib, afatinib, AZD9291, CO-1686,or WZ4002. The method comprises administering to a subject in needthereof an effective amount of a compound of Formula I, or apharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof. In another aspect, the method further comprises administeringto the subject a second agent that prevents EGFR dimer formation.

Another aspect of the present application relates to a method oftreating or preventing cancer, wherein the cell of the cancer comprisesan activated EGFR. The method comprises administering to a subject inneed thereof an effective amount of a compound of Formula I, or apharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof. In another aspect, the method further comprises administeringto the subject a second agent that prevents EGFR dimer formation.

Another aspect of the present application relates to a method oftreating or preventing cancer in a subject, wherein the subject isidentified as being in need of EGFR inhibition for the treatment orprevention of cancer. The method comprises administering to the subjectan effective amount of a compound of Formula I, or a pharmaceuticallyacceptable salt, hydrate, solvate, or stereoisomer thereof. In anotheraspect, the method further comprises administering to the subject asecond agent that prevents EGFR dimer formation.

Another aspect of the present application relates to a method oftreating or preventing cancer, wherein the cell of the cancer comprisesan activated ERBB2. The method comprises administering to a subject inneed thereof an effective amount of a compound of Formula or apharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof. In another aspect, the method further comprises administeringto the subject a second agent that prevents ERBB2 dimer formation.

Another aspect of the present application relates to a method oftreating or preventing cancer in a subject, wherein the subject isidentified as being in need of ERBB2 inhibition for the treatment orprevention of cancer. The method comprises administering to the subjectan effective amount of a compound of Formula I, or a pharmaceuticallyacceptable salt, hydrate, solvate, or stereoisomer thereof. In anotheraspect, the method further comprises administering to the subject asecond agent that prevents ERBB2 dimer formation.

Another aspect of the present application relates to a kit comprising acompound of Formula I, or a pharmaceutically acceptable salt, hydrate,solvate, or stereoisomer thereof. In another aspect, the kit furthercomprises a second agent that prevents EGFR dimer formation.

Another aspect of the present application relates to a compound ofFormula I, or a pharmaceutically acceptable salt, hydrate, solvate, orstereoisomer thereof, for use in the manufacture of a medicament for

inhibiting a kinase (e.g., EGFR)) in a subject in need thereof,

treating or preventing a disease (e.g., a disease in which EGFR plays arole) in a subject in need thereof,

treating or preventing a disease resistant to an EGER targeted therapy,such as a therapy with gefitinib, erlotinib, afatinib, AZD9291, CO-1686,or WZ4002, in a subject in need thereof,

treating or preventing cancer in a subject in need thereof, wherein thecell of the cancer comprises an activated EGFR or an activated ERBB2, or

treating or preventing cancer in a subject, wherein the subject isidentified as being in need of EGFR inhibition or ERBB2 inhibition forthe treatment or prevention of cancer.

In another aspect, the present application relates to a compound ofFormula I, or a pharmaceutically acceptable salt, hydrate, solvate, orstereoisomer thereof, and a second agent that prevents EGFR dimerformation, for use in the manufacture of a medicament for

inhibiting a kinase (e.g., EGFR) in a subject in need thereof,

treating or preventing a disease (e.g., a disease in which EGFR plays arole) in a subject in need thereof,

treating or preventing a disease resistant to an EGFR targeted therapy,such as a therapy with gefitinib, erlotinib, afatinib, AZD9291, CO-1686,or WZ4002, in a subject in need thereof,

treating or preventing cancer in a subject in need thereof, wherein thecell of the cancer comprises an activated EGFR or an activated ERBB2, or

treating or preventing cancer in a subject, wherein the subject isidentified as being in need of EGFR inhibition or ERBB2 inhibition forthe treatment or prevention of cancer.

Another aspect of the present application relates to a compound ofFormula I, or a pharmaceutically acceptable salt, hydrate, solvate, orstereoisomer thereof, for

inhibiting a kinase e.g. EGFR) in a subject in need thereof,

treating or preventing a disease (e.g., a disease in which EGFR plays arole) in a subject need thereof,

treating or preventing a disease resistant to an EGFR targeted therapy,such as a therapy with gefitinib, erlotinib, afatinib, AZD9291, CO-1686,or WZ4002, in a subject in need thereof,

treating or preventing cancer in a subject in need thereof, wherein thecell of the cancer comprises an activated EGFR or an activated ERBB2, or

treating or preventing cancer in a subject, wherein the subject isidentified as being in need of EGFR inhibition or ERBB2 inhibition forthe treatment or prevention of cancer.

Another aspect of the present application relates to a compound ofFormula I, or a pharmaceutically acceptable salt, hydrate, solvate, orstereoisomer, stereoisomer thereof, and a second agent that preventsEGFR dimer formation, for

inhibiting a kinase (e.g., EGFR) in a subject in need thereof,

treating or preventing a disease e.g., a disease in which EGFR plays arole) in a subject need thereof,

treating or preventing a disease resistant to an EGFR targeted therapy,such as a therapy with gefitinib, erlotinib, afatinib, AZD9291, CO-1686,or WZ4002, in a subject in need thereof,

treating or preventing cancer in a subject in need thereof, wherein thecell of the cancer comprises an activated EGFR or an activated ERBB2, or

treating or preventing cancer in a subject, wherein the subject isidentified as being in need of EGFR inhibition or ERBB2 inhibition forthe treatment or prevention of cancer.

Another aspect of the present application relates to use of a compoundof Formula I, or a pharmaceutically acceptable salt, hydrate, solvate,or stereoisomer thereof, in the manufacture of a medicament for

inhibiting a kinase (e.g., EGFR) in a subject in need thereof,

treating or preventing a disease (e.g., a disease in which EGFR plays arole) in a subject in need thereof,

treating or preventing a disease resistant to an EGFR targeted therapy,such as a therapy with gefitinib, erlotinib, afatinib, AZD9291, CO-1686,or WZ4002, in a subject in need thereof,

treating or preventing cancer in a subject in need thereof, wherein thecell of the cancer comprises an activated EGFR or an activated ERBB2, or

treating or preventing cancer in a subject, wherein the subject isidentified as being in need of EGFR inhibition or ERBB2 inhibition forthe treatment or prevention of cancer.

In another aspect, the present application relates to use of a compoundof Formula I, or a pharmaceutically acceptable salt, hydrate, solvate,or stereoisomer thereof, and a second agent that prevents EGFR dimerformation, in the manufacture of a medicament for

inhibiting a kinase (e.g., EGFR) in a subject in need thereof,

treating or preventing a disease (e.g., a disease in which EGFR plays arole) in a subject in need thereof,

treating or preventing a disease resistant to an EGFR targeted therapy,such as a therapy with gefitinib, erlotinib, afatinib, AZD9291, CO-1686,or WZ4002, in a subject in need thereof,

treating or preventing cancer in a subject in need thereof, wherein thecell of the cancer comprises an activated. EGFR or an activated ERBB2,or

treating or preventing cancer in a subject, wherein the subject isidentified as being in need of EGFR inhibition or ERBB2 inhibition forthe treatment or prevention of cancer.

Another aspect of the present application relates to use of a compoundof Formula I, or a pharmaceutically acceptable salt, hydrate, solvate,or stereoisomer thereof, in

inhibiting a kinase (e.g., EGFR) in a subject in need thereof,

treating or preventing a disease (e.g., a disease in which EGFR plays arole) in a subject in need thereof,

treating or preventing a disease resistant to an EGFR targeted therapy,such as a therapy with gefitinib, erlotinib, afatinib, AZD9291, CO-1686,or WZ4002, in a subject in need thereof,

treating or preventing cancer in a subject in need thereof, wherein thecell of the cancer comprises an activated EGFR or an activated ERBB2, or

treating or preventing cancer in a subject, wherein the subject isidentified as being in need of EGFR inhibition or ERBB2 inhibition forthe treatment or prevention of cancer.

Another aspect of the present application relates to use of a compoundof Formula I, or a pharmaceutically acceptable salt, hydrate, solvate,or stereoisomer thereof, and a second agent that prevents EGFR dimerformation, in

inhibiting a kinase (e.g., EGFR) in a subject in need thereof,

treating or preventing a disease (e.g., a disease in which EGFR plays arole) in a subject in need thereof,

treating or preventing a disease resistant to an EGFR targeted therapy,such as a therapy with gefitinib, erlotinib, afatinib, AZD9291, CO-1686,or WZ4002, in a subject in need thereof,

treating or preventing cancer in a subject in need thereof, wherein thecell of the cancer comprises an activated EGFR or an activated ERBB2, or

treating or preventing cancer in a subject, wherein the subject isidentified as being in need of EGFR inhibition or ERBB2 inhibition forthe treatment or prevention of cancer.

The present application provides inhibitors of EGFR, such as EGFRcontaining one or more mutations, that are therapeutic agents in thetreatment or prevention of diseases such as cancer and metastasis.

The present application further provides compounds and compositions withan improved efficacy and/or safety profile relative to known EGFRinhibitors. The present application also provides agents with novelmechanisms of action toward. EGFR kinases in the treatment or preventionof various types of diseases including cancer and metastasis.

The details of the application are set forth in the accompanyingdescription below. Although methods and materials similar or equivalentto those described herein can be used in the practice or testing of thepresent application, illustrative methods and materials are nowdescribed. Other features, objects, and advantages of the applicationwill be apparent from the description and from the claims. In thespecification and the appended claims, the singular forms also includethe plural unless the context clearly dictates otherwise. Unless definedotherwise, all technical and scientific terms used herein have the samemeaning as commonly understood by one of ordinary skill in the art towhich this application belongs. The contents of all references(including literature references, issued patents, published patentapplications, and co-pending patent applications) cited throughout thisapplication are hereby expressly incorporated herein in their entiretiesby reference.

DETAILED DESCRIPTION Compounds of the Application

The compounds of the present application are benzodiazepine-basedcompounds that modulate EGFR allosterically. In one aspect, the presentapplication relates to a compound of Formula I:

or a pharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof, wherein:

A₁ is phenyl or 5- or 6-membered heteroaryl comprising 1-3 heteroatomsselected from N, O, and S, wherein the phenyl or heteroaryl isoptionally substituted with one or more R₀;

each R₀ is independently C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆hydroxyalkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkoxy, OH, halogen, CN, phenyl,C₃-C₆ cycloalkyl, 5- or 6-membered heteroaryl comprising 1-3 heteroatomsselected from N, O, and S, or 5- or 6-membered heterocyclyl comprising1-3 heteroatoms selected from N, O, and S, wherein the phenyl,cycloalkyl, heteroaryl, or heterocyclyl is optionally substituted withone or more substituents independently selected from C₁-C₆ alkyl, C₁-C₆haloalkyl, C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkoxy, OH, andhalogen, or

two R₀, taken together with the adjacent atoms to which they areattached, form phenyl, C₅-C₆ cycloalkyl, 5- or 6-membered heteroarylcomprising 1-3 heteroatoms selected from N, O, and S, or 5- or6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O,and S, wherein the phenyl, cycloalkyl, heteroaryl, or heterocyclyl isoptionally substituted with one or more substituents independentlyselected from C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆ hydroxyalkyl, C₁-C₆alkoxy, C₁-C₆ haloalkoxy, OH, and halogen;

R₁ is H, C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy,C₁-C₆ haloalkoxy, or (CH₂)_(m)-A₂;

A₂ is phenyl, C₃-C₆ cycloalkyl, 5- or 6-membered heteroaryl comprising1-3 heteroatoms selected from N, O, and S, or 5- or 6-memberedheterocyclyl comprising 1-3 heteroatoms selected from N, O, and S,wherein the phenyl, cycloalkyl, heteroaryl, or heterocyclyl isoptionally substituted with one or more substituents independentlyselected from C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆ hydroxyalkyl, C₁-C₆alkoxy, C₁-C₆ haloalkoxy, OH, and halogen;

m is 0, 1, 2, or 3;

R₂ is 11 or C₁-C₆ alkyl;

R₃ is H, halogen, C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆ hydroxyalkyl,C₁-C₆ alkoxy, or C₁-C₆ haloalkoxy; or

R₂ and R₃, taken together with the intervening atoms, form a 5- or6-membered heterocyclyl ring comprising 0-2 additional heteroatomsselected from N, O, and S, or a 5- or 6-membered heteroaryl ringcomprising 0-2 additional heteroatoms selected from N, O, and S, whereinthe heterocyclyl or heteroaryl is optionally substituted with one ormore substituents independently selected from oxo, halogen, C₁-C₆ alkyl,C₁-C₆ haloalkyl, C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy, and C₁-C₆ haloalkoxy;

L₁ is a bond, a straight or branched bivalent C₁-C₆ alkylene chain,—C(O)—, —C(O)(CH₂)_(n)—, —C(O)NH—, —C(O)NH(CH₂)_(n)—, —C(O)N(C₁-C₆alkyl)-, —C(O)N(C₁-C₆ alkyl)(CH₂)_(n)—, —C(O)O—, —C(O)O(CH₂)_(n)—,—C(S)—, —C(S)(CH₂)_(n)—, —C(S)NH—, —C(S)N(C₁-C₆ alkyl)-,—C(S)NH(CH₂)_(n)—, —C(S)N(C₁-C₆ alkyl)(CH₂)_(n)—, —(CH₂)_(p)NH,—(CH₂)_(p)N(C₁-C₆ alkyl)-, or —(CH₂)_(p)O—;

n is 1 or 2;

p is 1, 2, or 3

X₁, X₂, X₃, and X₄ are each independently N or CR₄, provided that atleast two of X₁, X₂, X₃, and X₄ are CR₄;

each R₄ is independently H, NR₅R₆, NR₇C(O)R₈, C₁-C₆ alkyl, C₁-C₆haloalkyl, C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkoxy, OH,halogen, CN, phenyl, C₃-C₆ cycloalkyl, 5- or 6-membered heteroarylcomprising 1-3 heteroatoms selected from N, O, and S, or 5- or6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O,and S, wherein the phenyl, cycloalkyl, heteroaryl, or heterocyclyl isoptionally substituted with one or more substituents independentlyselected from C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆ hydroxyalkyl, C₁-C₆alkoxy, C₁-C₆ haloalkoxy, OH, and halogen;

each R5 and each R6 are independently H or C₁-C₄ alkyl;

each R7 is independently H or C₁-C₄ alkyl;

each R8 is independently C₁-C₄ alkyl;

X₅, X₆, X₇, and X₈ are each independently N, CR₉, or CR₁₀, provided thatat least one of X₅, X₆, X₇, and X₈ is CR₉ and at least one of X₅, X₆,X₇, and X₈ is CR₁₀;

each R₉ is independently H, halogen, OH, CN, C₁-C₆ alkyl, C₁-C₆haloalkyl, C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy, or C₁-C₆ haloalkoxy;

each R₁₀ is independently phenyl, C₃-C₆ cycloalkyl, 5- or 6-memberedheteroaryl comprising 1-4 heteroatoms selected from N, O, and S, or 5-or 6-membered heterocyclyl comprising 1-3 heteroatoms selected from N,O, and S, wherein the phenyl, cycloalkyl, heteroaryl, or heterocyclyl isoptionally substituted with one or more R₁₁;

each R₁₁ is independently C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆hydroxyalkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkoxy, OH, halogen, CN, phenyl,C₃-C₆ cycloalkyl, 5- or 6-membered heteroaryl comprising 1-3 heteroatomsselected from N, O, and S, or 5- or 6-membered heterocyclyl comprising1-3 heteroatoms selected from N, O, and S, wherein the phenyl,cycloalkyl, heteroaryl, or heterocyclyl is optionally substituted withone or more substituents independently selected from C₁-C₆ alkyl, C₁-C₆haloalkyl, C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy. C₁-C₆ haloalkoxy, OH, andhalogen, or

two R₁₁, taken together with the adjacent atoms to which they areattached, form phenyl, C₅-C₆ cycloalkyl, 5- or 6-membered heteroarylcomprising 1-3 heteroatoms selected from N, O, and S, or 5- or6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O,and S, wherein the phenyl, cycloalkyl, heteroaryl, or heterocyclyl isoptionally substituted with one or more substituents independentlyselected from C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆ hydroxyalkyl, C₁-C₆alkoxy, C₁-C₆ haloalkoxy, OH, and halogen.

For a compound of Formula I, where applicable, each of the variables canbe a group as described below.

(I1) In one embodiment, A₁ is phenyl.

(I2) In one embodiment, A₁ is 5- or 6-membered heteroaryl comprising 1-3heteroatoms selected from N, O, and S.

(I3) In one embodiment, A₁ is 5-membered heteroaryl comprising 1-3heteroatoms selected from N, O, and S. In one embodiment, A₁ is5-membered heteroaryl comprising 1-2 heteroatoms selected from N, O, andS. In one embodiment, A₁ is 5-membered heteroaryl comprising 1-2heteroatoms selected from N and O. In one embodiment, A₁ is heteroarylselected from pyrrolyl, furanyl, thiophenyl, pyrazolyl, imidazolyl,oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, triazolyl, oxadiazolyl,and thiadiazolyl. In one embodiment, A₁ is pyrazolyl or imidazolyl.

(I4) In one embodiment, A₁ is 6-membered heteroaryl comprising 1-3heteroatoms selected from N, O, and S. In one embodiment, A₁ is6-membered heteroaryl comprising 1-2 heteroatoms selected from N, O, andS. In one embodiment, A₁ is 6-membered heteroaryl comprising 1-2heteroatoms selected from N and O. In one embodiment, A₁ is heteroarylselected from pyridinyl, pyridazinyl, pyrimidinyl pyrazinyl, pyranyl,thiopyranyl, thiazinyl, dioxinyl, and triazinyl. In one embodiment, A₁is 6-membered heteroaryl comprising 1-2 nitrogen atoms. In oneembodiment, A₁ is 6-membered heteroaryl comprising 1 nitrogen atom. Inone embodiment, A₁ is 6-membered heteroaryl comprising 2 nitrogen atoms.In one embodiment, A₁ is heteroaryl selected from pyridinyl,pyridazinyl, and pyrimidinyl. In one embodiment, A₁ is pyridinyl. In oneembodiment, A₁ is pyridazinyl. In one embodiment, A₁ is pyrimidinyl.

(II1) In one embodiment, each R₀ is independently C₁-C₆ straight-chainor C₃-C₆ branched alkyl (e.g., methyl, ethyl, n-propyl, i-propyl,n-butyl; i-butyl, s-butyl, t-butyl, pentyl, or hexyl), C₁-C₆straight-chain or C₃-C₆ branched haloalkyl (e.g., methyl, ethyl,n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, pentyl, orhexyl, each of which is substituted with one or more halogen (e.g., F,Cl, Br, or I)), C₁-C₆ straight-chain or C₃-C₆ branched hydroxyalkyl(e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl,t-butyl, pentyl, or hexyl, each of which is substituted with one or moreOH), C₁-C₆ straight-chain or C₃-C₆ branched alkoxy (e.g., methoxy,ethoxy, n-propoxy, i-propoxy, n-butoxy, butoxy, s-butoxy, t-butoxy,pentoxy, or hexyloxy), C₁-C₆ straight-chain or C₃-C₆ branched haloalkoxyethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy, t-butoxy,pentoxy, or hexyloxy, each of which is substituted with one or morehalogen (e.g., F, Cl, Br, or I)), OH, halogen (e.g., F, Cl, Br, or I),CN, phenyl, C₃-C₆ cycloalkyl, 5- or 6-membered heteroaryl comprising 1-3heteroatoms selected from N, O, and S, or 5- or 6-membered heterocyclylcomprising 1-3 heteroatoms selected from N, O, and S, wherein thephenyl, cycloalkyl, heteroaryl, or heterocyclyl is optionallysubstituted with one or more substituents independently selected fromC₁-C₆ straight-chain or C₃-C₆ branched alkyl (e.g., methyl, ethyl,n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, pentyl, orhexyl), C₁-C₆ straight-chain or C₃-C₆ branched haloalkyl (e.g., methyl,ethyl, n-propyl, i-propyl, n-butyl, butyl, s-butyl, t-butyl, pentyl, orhexyl, each of which is substituted with one or more halogen (e.g., F,Cl, Br, or I)), C₁-C₆ straight-chain or C₃-C₆ branched hydroxyalkyl(e.g., methyl, ethyl, n-propyl, n-butyl, i-butyl, s-butyl, t-butyl,pentyl, or hexyl, each of which is substituted with one or more OH).C₁-C₆ straight-chain or C₃-C₆ branched alkoxy (e.g., methoxy, ethoxy,n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy, t-butoxy, pentoxy,or hexyloxy), C₁-C₆ straight-chain or C₃-C₆ branched haloalkoxy methoxy,ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy, t-butoxy,pentoxy, or hexyloxy, each of which is substituted with one or morehalogen e.g.; F, Cl, Br, or I)), OH, and halogen (e.g., F, Cl, Br, orI).

(II2) In one embodiment, each R₀ is independently C₁-C₄ straight-chainor C₃-C₄ branched alkyl (e.g., methyl, ethyl, n-propyl, i-propyl,n-butyl, i-butyl, s-butyl, ort-butyl), C₁-C₄ straight-chain or C₃-C₄branched haloalkyl (e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl,i-butyl, s-butyl, or t-butyl, each of which is substituted with one ormore halogen (e.g., F, Cl, Br, or I)), C₁-C₄ straight-chain or C₃-C₄branched hydroxyalkyl (e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl,i-butyl, s-butyl, or t-butyl, each of which is substituted with one ormore OH), C₁-C₄ straight-chain or C₃-C₄ branched alkoxy (e.g., methoxy,ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy, ort-butoxy), C₁-C₄ straight-chain or C₃-C₄ branched haloalkoxy (e.g.,methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy, ort-butoxy, each of which is substituted with one or more halogen (e.g.,F, Cl, Br, or I)), OH, halogen (e.g., F, Cl, Br, or I), CN, phenyl,C₃-C₆ cycloalkyl, heteroaryl comprising one 5- or 6-membered ring and1-3 heteroatoms selected from N, O, and S, or heterocyclyl comprisingone 5- or 6-membered ring and 1-3 heteroatoms selected from N, O, and S,wherein the phenyl, cycloalkyl, heteroaryl, or heterocyclyl isoptionally substituted as described herein (e.g., as in (II1)).

(II3) In one embodiment, one or more R₀ is C₁-C₆ alkyl, e.g., methyl,ethyl, n-propyl, n-butyl, s-butyl, t-butyl, pentyl, or hexyl. In oneembodiment, one or more R_(D) is methyl.

(II4) In one embodiment, one or more R₀ is C₁-C₆ haloalkyl, e.g.,methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl,pentyl, or hexyl, each of which is substituted with one or more halogen(e.g., F, Cl, Br, or I). In one embodiment, one or more R₀ is CF₃, CHF₂,or CH₂F.

(II5) In one embodiment, one or more R₀ is C₁-C₆ hydroxyalkyl, e.g.,methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl,pentyl, or hexyl, each of which is substituted with one or more OH. Inone embodiment, one or more R₀ is CH₂OH, CH₂CH₂OH, or C(CH₃)₂OH.

(II6) In one embodiment, one or more R, is C₁-C₆ alkoxy, e.g., methoxy,ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy, t-butoxy,pentoxy, or hexyloxy. In one embodiment, one or more R₀ is methoxy.

(II7) In one embodiment, one or more R₀ is C₁-C₆ haloalkoxy, e.g.,methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy,t-butoxy, pentoxy, or hexyloxy, each of which is substituted with one ormore halogen (e.g., F, Cl, Br, or I). In one embodiment, one or more R₀is OCF₃, OCHF₂, or OCH₂F.

(II8) In one embodiment, one or more is OH.

(II9) In one embodiment, one or more R₀ is halogen, e.g., F, Br, Cl, orI.

(II10) In one embodiment, one or more R₀ is CN.

(II11) In one embodiment, one or more R₀ is phenyl, C₃-C₆ cycloalkyl, 5-or 6-membered heteroaryl comprising 1-3 heteroatoms selected from N, O,and S, or 5- or 6-membered heterocyclyl comprising 1-3 heteroatomsselected from N, O, and S, wherein the phenyl, cycloalkyl, heteroaryl,or heterocyclyl is optionally substituted as described herein (e.g., asin (II1)).

(II12) In one embodiment, one or more R₀ is phenyl, optionallysubstituted as described herein (e.g., as in (II1)).

(II13) In one embodiment, one or more R₀ is C₃-C₆ cycloalkyl, optionallysubstituted as described herein (e.g., as in (Iii)). In one embodiment,one or more R₀ is cyclopropyl. In one embodiment, one or more R₀ iscyclobutyl. In one embodiment, one or more R₀ is cyclopentyl. In oneembodiment, one or more R₀ is cyclohexyl.

(II14) In one embodiment, one or more R₀ is 5- or 6-membered heteroarylcomprising 1-3 heteroatoms selected from N, O, and S, optionallysubstituted as described herein (e.g., as in (II1).

(II15) In one embodiment, one or more R₀ is 5-membered heteroarylcomprising 1-3 heteroatoms selected from N, O, and S, optionallysubstituted as described herein (e.g., as in (II1)). In one embodiment,one or more R₀ is 5-membered heteroaryl comprising 1-2 heteroatomsselected from N, O, and S. In one embodiment, one or more R₀ is5-membered heteroaryl comprising 1-2 heteroatoms selected from N and O.In one embodiment, one or more R₀ is heteroaryl selected from pyrrolyl,furanyl, thiophenyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl,thiazolyl, isothiazolyl, triazolyl, oxadiazolyl, and thiadiazolyl, eachof which is optionally substituted as described herein (e.g., as in(II1)). In one embodiment, one or more R₀ is pyrazolyl or imidazolyl,optionally substituted as described herein (e.g., as in (II1)).

(II16) In one embodiment, one or more R₀ is 6-membered heteroarylcomprising 1-3 heteroatoms selected from N, O, and S, optionallysubstituted as described herein (e.g., as in (II1)). In one embodiment,one or more R₀ is 6-membered heteroaryl comprising 1-2 heteroatomsselected from N, O, and S. In one embodiment, one or more R₀ is6-membered heteroaryl comprising 1-2 heteroatoms selected from N and O.In one embodiment, one or more R₀ is heteroaryl selected from pyridinyl,pyridazinyl, pyrimidinyl, pyrazinyl, pyranyl, thiopyranyl, diazinyl,thiazinyl, dioxinyl, and triazinyl, each of which is optionallysubstituted as described herein (e.g., as in (II1)). In one embodiment,one or more R₀ is 6-membered heteroaryl comprising 1-2 nitrogen atoms.In one embodiment, one or more R₀ is 6-membered heteroaryl comprising 1nitrogen atom. In one embodiment, one or more R₀ is 6-memberedheteroaryl comprising 2 nitrogen atoms. In one embodiment, one or moreR₀ is heteroaryl selected from pyridinyl, pyridazinyl, and pyrimidinyl,each of which is optionally substituted as described herein (e.g., as in(II1)).

(II17) In one embodiment, one or more R₀ is 5- or 6-memberedheterocyclyl comprising 1-3 heteroatoms selected from N, O, and S,optionally substituted as described herein (e.g., as in (II1)).

(II18) In one embodiment, one or more R₀ is 5-membered heterocyclylcomprising 1-3 heteroatoms selected from N, O, and S, optionallysubstituted as described herein (e.g., as in (II1)). In one embodiment,one or more R₀ is 5-membered heterocyclyl comprising 1-2 heteroatomsselected from N, O, and S. In one embodiment, one or more R₀ is5-membered heterocyclyl comprising 1-2 heteroatoms selected from N andO. In one embodiment, one or more R₀ is 5-membered heterocyclylcomprising 1 heteroatom selected from N, O, and S. In one embodiment,one or more R₀ is heterocyclyl selected from pyrrolidinyl,tetrahydrofuranyl, tetrahydrothiophenyl, pyrazolidinyl, imidazolidinyl,oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolidinyl,triazolidinyl, oxadiazolidinyl, isoxadiazolidinyl, thiadiazolidinyl, andisothiadiazolidinyl, each of which is optionally substituted asdescribed herein (e.g., as in (Ill)).

(II19) In one embodiment, one or more R₀ is 6-membered heterocyclylcomprising 1-3 heteroatoms selected from N, O, and S, optionallysubstituted as described herein (e.g., as in (II1)). In one embodiment,one or more R₀ is 6-membered heterocyclyl comprising 1-2 heteroatomsselected from N, O, and S. In one embodiment, one or more R₀ is6-membered heterocyclyl comprising 1-2 heteroatoms selected from N andO. In one embodiment, one or more R₀ is 6-membered heterocyclylcomprising 1 heteroatom selected from N, O, and S. In one embodiment,one or more R₀ is heterocyclyl selected from piperidinyl, piperazinyl,tetrahydropyranyl, hexahydropyridazinyl, hexahydropyrimidinyl,morpholinyl, and triazinanyl, each of which is optionally substituted asdescribed herein (e.g., as in (II1)). In one embodiment, one or more R₀is piperidinyl or piperazinyl, each of which is optionally substitutedas described herein (e.g., as in (II1)).

(II20) In one embodiment, two R₀, taken together with the adjacent atomsto which they are attached, form phenyl, C₅-C₆ cycloalkyl, 5- or6-membered heteroaryl comprising 1-3 heteroatoms selected from N, O, andS, or 5- or 6-membered heterocyclyl comprising 1-3 heteroatoms selectedfrom N, O, and S, wherein the phenyl, cycloalkyl, heteroaryl, orheterocyclyl is optionally substituted with one or more substituentsindependently selected from C₁-C₆ straight-chain or C₃-C₆ branched alkyl(e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl,t-butyl, pentyl, or hexyl), C₁-C₆ straight-chain or C₃-C₆ branchedhaloalkyl (e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl,s-butyl, t-butyl, pentyl, or hexyl, each of which is substituted withone or more halogen (e.g., F, Cl, Br, or I)). C₁-C₆ straight-chain orC₃-C₆ branched hydroxyalkyl (e.g., methyl, ethyl, n-propyl, i-propyl,n-butyl, i-butyl, s-butyl, t-butyl, pentyl, or hexyl, each of which issubstituted with one or more OH), C₁-C₆ straight-chain or C₃-C₆ branchedalkoxy (e.g., methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy,s-butoxy, t-butoxy, pentoxy, or hexyloxy), C₁-C₆ straight-chain or C₃-C₆branched haloalkoxy (e.g., methoxy, ethoxy, n-propoxy, i-propoxy,n-butoxy, i-butoxy, s-butoxy, t-butoxy, pentoxy, or hexyloxy, each ofwhich is substituted with one or more halogen (e.g., F, Cl, Br, or I)),OH, and halogen (e.g., F, Cl, Br, or I).

(II21) In one embodiment, two R₀, taken together with the adjacent atomsto which they are attached, form phenyl, optionally substituted asdescribed herein (e.g., as in (II20)). In one embodiment, A₁ takentogether with two R₀ forms naphthalenyl.

(II22) In one embodiment, two R₀, taken together with the adjacent atomsto which they are attached, form C₅-C₆ cycloalkyl, optionallysubstituted as described herein (e.g., as in (II20)). In one embodiment,A₁ taken together with two R₀ forms a ring system selected from indenyl,dihydroindenyl, tetrahydronaphthalenyl, and dihydronaphthalenyl.

(II23) In one embodiment, two R₀, taken together with the adjacent atomsto which they are attached, form 5- or 6-membered heterocyclylcomprising 1-3 heteroatoms selected from N, O, and S, optionallysubstituted as described herein (e.g., as in (II20)). In one embodiment,two R₀, taken together with the adjacent atoms to which they areattached, form 5-membered heterocyclyl comprising 1-2 heteroatomsselected from N, O, and S. In one embodiment, two R₀, taken togetherwith the adjacent atoms to which they are attached, form 5-memberedheterocycle comprising 1 heteroatom selected from N, O, and S. In oneembodiment, two R₀, taken together with the adjacent atoms to which theyare attached, form 6-membered heterocyclyl comprising 1-2 heteroatomsselected from N, O, and S. In one embodiment, two R₀, taken togetherwith the adjacent atoms to which they are attached, form 6-memberedheterocycle comprising 1 heteroatom selected from N, O, and S. In oneembodiment, A₁ taken together with two R₀ forms a ring system selectedfrom indolinyl, oxindolyl, isoindolinyl, dihydroindazolyl,dihydrobenzoimidazolonyl, dihydrobenzofuryl, benzofuranonyl,dihydrobenzothiophenyl, tetrahydroquinolinyl, dihydroquinolinonyl,tetrahydroquinoxazolinyl, dihydrobenzooxazinyl benzooxazolonyl,benzodioxolonyl, chromanyl, chromanonyl, dihydrobenzodioxinyl,thiochromanyl, dihydrobenzooxathiinyl, dihydrobenzodithiinyl, anddihydrobenzothiazinyl.

(II24) In one embodiment, two R₀, taken together with the adjacent atomsto which they are attached, form 5- or 6-membered heteroaryl comprising1-3 heteroatoms selected from N, O, and S, optionally substituted asdescribed herein (e.g., as in (II20)). In one embodiment, two R₀, takentogether with the adjacent atoms to which they are attached, form5-membered heteroaryl comprising 1-2 heteroatoms selected from N, O, andS. In one embodiment, two R₀, taken together with the adjacent atoms towhich they are attached, form 5-membered heteroaryl comprising 1-2heteroatoms selected from N and O. In one embodiment, two R₀, takentogether with the adjacent atoms to which they are attached, form5-membered heteroaryl comprising 1 heteroatom selected from N, O, and S.In one embodiment, A₁ taken together with two R₀ forms a ring systemselected from indolyl, indazolyl, benzoimidazolyl, benzotriazolyl,benzooxadiazolyl, benzooxazolyl, benzoisoxazolyl, benzofuryl,benzothiophenyl, benzothiazolyl, benzoisothiazolyl, benzothiadiazolyl,pyrrolopyridinyl, pyrrolopyrazinyl, pyrrolopyrimidinyl,imidazopyridinyl, purinyl, imidazopyrazinyl, pyrazolopyridinyl,pyrazolopyrimidinyl, pyrazolopyrazinyl, triazolopyridinyl,furopyridinyl, furopyrimidinyl, furopyrazinyl, thienopyridinyl,thicnopyrimidinyl, thienopyrazinyl, oxazolopyridinyl,oxazolopyrimidinyl, oxazolopyrazinyl, isoxazolopyridinyl,isoxazolopyrimidinyl, isoxazolopyrazinyl, thiazolopyridinyl,thiazolopyrimidinyl, thiazolopyrazinyl, and oxadiazolopyridinyl. In oneembodiment, two R₀, taken together with the adjacent atoms to which theyare attached, form 6-membered heteroaryl comprising 1-2 heteroatomsselected from N, O, and S. In one embodiment, two R₀, taken togetherwith the adjacent atoms to which they are attached, form 6-memberedheteroaryl comprising 1-2 heteroatoms selected from N and O. In oneembodiment, two R₀, taken together with the adjacent atoms to which theyare attached, form 6-membered heteroaryl comprising 1 heteroatomselected from N, O, and S. In one embodiment, two R₀, taken togetherwith the adjacent atoms to which they are attached, form 6-memberedheteroaryl comprising 1-3 nitrogen atoms. In one embodiment, two R₀,taken together with the adjacent atoms to which they are attached, form6-membered heteroaryl comprising 1-2 nitrogen atoms. In one embodiment,two R₀, taken together with the adjacent atoms to which they areattached, form 6-membered heteroaryl comprising 1 nitrogen atom. In oneembodiment, A₁ taken together with two R₀ forms a ring system selectedfrom quinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl, cinnolinyl,phthalazinyl, benzotriazinyl, naphthyridinyl, pyridopyrimidinyl,pteridinyl, pyridopyridazinyl, and pyrazinopyridazinyl.

(III1) In one embodiment, R₁ is H, C₁-C₆ straight-chain or C₃-C₆branched alkyl (e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl,i-butyl, s-butyl, t-butyl, pentyl, or hexyl), C₁-C₆ straight-chain orC₃-C₆ branched haloalkyl (e.g., methyl, ethyl, n-propyl, i-propyl,n-butyl, i-butyl, s-butyl, t-butyl, pentyl, or hexyl, each of which issubstituted with one or more halogen (e.g., F, Cl, Br, or I)), C₁-C₆straight-chain or C₃-C₆ branched hydroxyalkyl (e.g., methyl, ethyl,n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, pentyl, orhexyl, each of which is substituted with one or more OH), C₁-C₆straight-chain or C₃-C₆ branched alkoxy (e.g., methoxy, ethoxy,n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy, t-butoxy, pentoxy,or hexyloxy), or C₁-C₆ straight-chain or C₃-C₆ branched haloalkoxy(e.g., methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy,s-butoxy, t-butoxy, pentoxy, or hexyloxy, each of which is substitutedwith one or more halogen (e.g., F, Cl, Br, or I)).

(III2) In one embodiment, R₁ is H, C₁-C₆ alkyl, C₁-C₆ haloalkyl, orC₁-C₆ hydroxyalkyl, as described herein (e.g., as in (III1)).

(III3) In one embodiment, R₁ is H.

(III4) In one embodiment, R₁ is C₁-C₆ alkyl, e.g., methyl, ethyl,n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, pentyl, orhexyl. In one embodiment, R₁ is methyl.

(III5) In one embodiment, R₁ is C₁-C₆ haloalkyl, e.g., methyl, ethyl,n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, pentyl, orhexyl, each of which is substituted with one or more halogen (e.g., F,Cl, Br, or I). In one embodiment, R₁ is CF₃, CHF₂, or CH₂F.

(III6) In one embodiment, R₁ is C₁-C₆ hydroxyalkyl, e.g., methyl, ethyl,n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, pentyl, orhexyl, each of which is substituted with one or more OH. In oneembodiment. R₁ is CH₂OH, CH₂CH₂OH, or C(CH₃)₂OH.

(III7) In one embodiment, R₁ is C₁-C₆ alkoxy, e.g., methoxy, ethoxy,n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy, t-butoxy, pentoxy,or hexyloxy. In one embodiment, R₁ is methoxy.

(III8) In one embodiment, R₁ is C₁-C₆ haloalkoxy, e.g., methoxy, ethoxy,n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy, t-butoxy, pentoxy,or hexyloxy, each of which is substituted with one or more halogen(e.g., F, Cl, Br, or I).

(III9) In one embodiment, R₁ is (CH₂)_(m)-A₂.

(III10) In one embodiment, R₁ is (CH₂)₃-A₂.

(III11) In one embodiment, R₁ is (CH₂)₂-A₂.

(III12) In one embodiment, R₁ is (CH₂)₁-A₂.

(III13) In one embodiment, R₁ is (CH₂)₀-A₂.

(IV1) In one embodiment, A₂ is phenyl, C₃-C₆ cycloalkyl, 5- or6-membered heteroaryl comprising 1-3 heteroatoms selected from N, O, andS, or 5- or 6-membered heterocyclyl comprising 1-3 heteroatoms selectedfrom N, O, and S, wherein the phenyl, cycloalkyl, heteroaryl, orheterocyclyl is optionally substituted with one or more substituentsindependently selected from C₁-C₆ straight-chain or C₃-C₆ branched alkyl(e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl,t-butyl, pentyl, or hexyl), C₁-C₆ straight-chain or C₃-C₆ branchedhaloalkyl (e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl,s-butyl, t-butyl, pentyl, or hexyl, each of which is substituted withone or more halogen (e.g., F, Cl, Br, or I)), C₁-C₆ straight-chain orC₃-C₆ branched hydroxyalkyl (e.g., methyl, ethyl, n-propyl, i-propyl,n-butyl, i-butyl, s-butyl, t-butyl, pentyl, or hexyl, each of which issubstituted with one or more OH), C₁-C₆ straight-chain or C₃-C₆ branchedalkoxy (e.g., methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy,s-butoxy, t-butoxy, pentoxy, or hexyloxy), C₁-C₆ straight-chain or C₃-C₆branched haloalkoxy (e.g., methoxy, ethoxy, n-propoxy, i-propoxy,n-butoxy, i-butoxy, s-butoxy, t-butoxy, pentoxy, or hexyloxy, each ofwhich is substituted with one or more halogen (e.g., F, Cl, Br, or I)),OH, and halogen (e.g., F, Cl, Br, or I).

(IV2) In one embodiment, A₂ is phenyl, optionally substituted asdescribed herein (e.g., as in (IV1)).

(IV3) In one embodiment, A₂ is C₃-C₆ cycloalkyl, optionally substitutedas described herein (e.g., as in (IV1)). In one embodiment, A₂ iscyclopropyl. In one embodiment, A₂ is cyclobutyl. In one embodiment, A₂is cyclopentyl. In one embodiment, A₂ is cyclohexyl.

(IV4) In one embodiment, A₂ is 5- or 6-membered heteroaryl comprising1-3 heteroatoms selected from N, O, and S, optionally substituted asdescribed herein (e.g., as in (IV1)).

(IV5) In one embodiment, A₂ is 5-membered heteroaryl comprising 1-3heteroatoms selected from N, O, and S, optionally substituted asdescribed herein (e.g., as in (IV1)). In one embodiment, A₂ is5-membered heteroaryl comprising 1-2 heteroatoms selected from N, O, andS. In one embodiment. A₂ is 5-membered heteroaryl comprising 1-2heteroatoms selected from N and O. In one embodiment, A₂ is heteroarylselected from pyrrolyl, furanyl, thiophenyl, pyrazolyl, imidazolyl,oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, triazolyl, oxadiazolyl,and thiadiazolyl, each of which is optionally substituted as describedherein (e.g., as in (IV1)).

(IV6) In one embodiment, A₂ is 6-membered heteroaryl comprising 1-3heteroatoms selected from N, O, and S, optionally substituted asdescribed herein (e.g., as in (IV1)). In one embodiment, A₂ is6-membered heteroaryl comprising 1-2 heteroatoms selected from N, O, andS. In one embodiment, A₂ is 6-membered heteroaryl comprising 1-2heteroatoms selected from N and O. In one embodiment, A₂ is heteroarylselected from pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, pyranyl,thiopyranyl, diazinyl, thiazinyl, dioxinyl, and triazinyl, each of whichis optionally substituted as described herein (e.g., as in (IV1)). Inone embodiment, A₂ is 6-membered heteroaryl comprising 1-2 nitrogenatoms. In one embodiment, A₂ is membered heteroaryl comprising 1nitrogen atom. In one embodiment, A₂ is 6-membered heteroaryl comprising2 nitrogen atoms. In one embodiment, A₂ is heteroaryl selected frompyridinyl, pyridazinyl, and pyrimidinyl, each of which is optionallysubstituted as described herein (e.g., as in (IV1)).

(IV7) In one embodiment, A₂ is 5- or 6-membered heterocyclyl comprising1-3 heteroatoms selected from N, O, and S, optionally substituted asdescribed herein (e.g., as in (IV1)).

(IV8) In one embodiment, A₂ is 5-membered heterocyclyl comprising 1-3heteroatoms selected from N, O, and S, optionally substituted asdescribed herein (e.g., as in (IV1)). In one embodiment, A₂ is5-membered heterocyclyl comprising 1-2 heteroatoms selected from N, O,and S. In one embodiment, A₂ is 5-membered heterocyclyl comprising 1-2heteroatoms selected from N and O. In one embodiment, A₂ is 5-memberedheterocyclyl comprising 1 heteroatom selected from N, O, and S. In oneembodiment, A₂ is heterocyclyl selected from pyrrolidinyl,tetrahydrofuranyl, tetrahydrothiophenyl, pyrazolidinyl, imidazolidinyl,oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolidinyl,triazolidinyl, oxadiazolidinyl, isoxadiazolidinyl, thiadiazolidinyl, andisothiadiazolidinyl, each of which is optionally substituted asdescribed herein (e.g., as in (IV1)).

(IV9) In one embodiment, A₂ is 6-membered heterocyclyl comprising 1-3heteroatoms selected from N, O, and S, optionally substituted asdescribed herein (e.g., as in (IV1)). In one embodiment, A₂ is6-membered heterocyclyl comprising 1-2 heteroatoms selected from N, O,and S. In one embodiment, A₂ is 6-membered heterocyclyl comprising 1-2heteroatoms selected from N and O. In one embodiment, A₂ is 6-memberedheterocyclyl comprising 1 heteroatom selected from N, O, and S. In oneembodiment, A₂ is heterocyclyl selected from piperidinyl, piperazinyl,tetrahydropyranyl, hexahydropyridazinyl, hexahydropyrimidinyl,morpholinyl, and triazinanyl, each of which is optionally substituted asdescribed herein (e.g., as in (IV1)). In one embodiment, A₂ ispiperidinyl or piperazinyl, each of which is optionally substituted asdescribed herein (e.g., as in (IV1)).

(V1) In one embodiment, m is 0, 1, or 2.

(V2) In one embodiment, m is 0 or 1.

(V3) In one embodiment, m is 0.

(V4) In one embodiment, m is 1, 2, or 3.

(V5) In one embodiment, m is 1 or 2.

(V6) In one embodiment, m is 1.

(V7) In one embodiment, m is 2 or 3.

(V8) In one embodiment, m is 2.

(V9) In one embodiment, m is 3.

(VI1) In one embodiment, R₂ is H.

(VI2) In one embodiment, R₂ is C₁-C₆ straight-chain or C₃-C₆ branchedalkyl (e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl,s-butyl, t-butyl, pentyl, or hexyl). In one embodiment, R₂ is methyl.

(VII1) In one embodiment, R₃ is H, halogen (e.g., F, Cl, Br, or I),C₁-C₆ straight-chain or C₃-C₆ branched alkyl (e.g., methyl, ethyl,n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, pentyl, orhexyl), C₁-C₆ straight-chain or C₃-C₆ branched haloalkyl (e.g., methyl,ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, pentyl,or hexyl, each of which is substituted with one or more halogen (e.g.,F, Cl, Br, or I)), C₁-C₆ straight-chain or C₃-C₆ branched hydroxyalkyl(e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl,t-butyl, pentyl, or hexyl, each of which is substituted with one or moreOH), C₁-C₆ straight-chain or C₃-C₆ branched alkoxy (e.g., methoxy,ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy, t-butoxy,pentoxy, or hexyloxy), or C₁-C₆ straight-chain or C₃-C₆ branchedhaloalkoxy (e.g., methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy,i-butoxy, s-butoxy, t-butoxy, pentoxy, or hexyloxy, each of which issubstituted with one or more halogen (e.g., F, Cl, Br, or I)).

(VII2) In one embodiment, R₃ is H.

(VII3) In one embodiment, R₃ is halogen. In one embodiment, R₃ is F. Inone embodiment, R₃ is Br. In one embodiment, R₃ is Cl. In oneembodiment. R₃ is I.

(VII4) In one embodiment, R₃ is C₁-C₆ straight-chain or C₃-C₆ branchedalkyl (e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl,s-butyl, t-butyl, pentyl, or hexyl), C₁-C₆ straight-chain or C₃-C₆branched haloalkyl (e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl,i-butyl, s-butyl, t-butyl, pentyl, or hexyl, each of which issubstituted with one or more halogen (e.g., F, Cl, Br, or I)), or C₁-C₆straight-chain or C₃-C₆ branched hydroxyalkyl (e.g., methyl, ethyl,n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, pentyl, orhexyl, each of which is substituted with one or more OH).

(VII5) In one embodiment, R₃ is C₁-C₆ straight-chain or C₃-C₆ branchedalkoxy (e.g., methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy,s-butoxy, t-butoxy, pentoxy, or hexyloxy), or C₁-C₆ straight-chain orC₃-C₆ branched haloalkoxy (e.g., methoxy, ethoxy, n-propoxy, i-propoxy,n-butoxy, i-butoxy, s-butoxy, t-butoxy, pentoxy, or hexyloxy, each ofwhich is substituted with one or more halogen (e.g., F, Cl, Br, or I)).

(VII6) In one embodiment, R₂ and R₃, taken together with the interveningatoms, form a 5- or 6-membered heterocyclyl ring comprising 0-2additional heteroatoms selected from N, O, and S, optionally substitutedwith one or more substituents independently selected from oxo, halogen(e.g., F, Cl, Br, or I), C₁-C₆ straight-chain or C₃-C₆ branched alkyl(e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl,t-butyl, pentyl, or hexyl), C₁-C₆ straight-chain or C₃-C₆ branchedhaloalkyl (e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl,s-butyl, t-butyl, pentyl, or hexyl, each of which is substituted withone or more halogen (e.g., F, Cl, Br, or I)), C₁-C₆ straight-chain orC₃-C₆ branched hydroxyalkyl (e.g., methyl, ethyl, n-propyl, i-propyl,n-butyl, i-butyl, s-butyl, t-butyl, pentyl, or hexyl, each of which issubstituted with one or more OH), C₁-C₆ straight-chain or C₃-C₆ branchedalkoxy (e.g., methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy,s-butoxy, t-butoxy, pentoxy, or hexyloxy), or C₁-C₆ straight-chain orC₃-C₆ branched haloalkoxy (e.g., methoxy, ethoxy, n-propoxy, i-propoxy,n-butoxy, i-butoxy, s-butoxy, t-butoxy, pentoxy, or hexyloxy, each ofwhich is substituted with one or more halogen (e.g., F, Cl, Br, or I)).In one embodiment, R₂ and R₃, taken together with the intervening atoms,form a heterocyclyl ring selected from pyrrolidinyl, imidazolinyl,oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolidinyl,tetrahydrofuranyl, dioxolanyl, tetrahydrothiophenyl, dithiolanyl,dihydropyridinyl, tetrahydropyridinyl, piperidinyl, dihydropyrazinyl,tetrahydropyrazinyl, piperazinyl, morpholinyl, oxazinyl,dihydrooxazinyl, oxazinanyl, dihydrothiazinyl, tetrahydrothiazinyl,thiomorpholinyl, pyranyl, dihydropyaanyl, tetrahydropyranyl, dioxinyl,dihydrodioxinyl, dioxanyl, thiopyranyl, dihydrothiopyranyl,tetrahydropyranyl, and dithianyl, each of which is optionallysubstituted as described herein.

(VII7) In one embodiment, R₂ and R₃, taken together with the interveningatoms, form a 5- or 6-membered heterocyclyl ring comprising 0-2additional heteroatoms selected from N, O, and S, wherein theheterocyclyl is substituted with oxo. In one embodiment, R₂ and R₃,taken together with the intervening atoms, form a heterocyclyl ringselected from pyrrolidin-2-onyl, pyrrolidin-3-onyl,pyrrolidine-2,3-dionyl pyrrolidine-2,5-dionyl, and imidazolidinonyl,oxazolidinonyl, and thiazolidinonyl. In one embodiment, the heterocycleis selected from piperidin-2-onyl, piperidin-3-onyl, piperidin-4-onyl,piperidine-2,6-dionyl, dihydropyrimidinonyl, tetrahydropyrimidinonyl,dihydropyridazinoyl, tetrahydropyridazinonyl, dihydrooxazinonyl,oxazinanonyl, thiazinanonyl, pyridinonyl, and dihydropyridinonyl.

(VII8) In one embodiment, R₂ and R₃, taken together with the interveningatoms, form a 5- or 6-membered heteroaryl ring comprising 0-2 additionalheteroatoms selected from N, O, and S, optionally substituted asdescribed herein (e.g., as in (VII6)). In one embodiment, R₂ and R₃,taken together with the intervening atoms, form a heteroaryl selectedfrom pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, pyranyl,thiopyranyl, diazinyl, thiazinyl, dioxinyl, and triazinyl, each of whichis optionally substituted as described herein (e.g., as in (VII6)). Inone embodiment, R₂ and R₃, taken together with the intervening atoms,form a 5-membered heteroaryl ring comprising 0-2 additional nitrogenatoms. In one embodiment, R₂ and R₃, taken together with the interveningatoms, form a heteroaryl selected from pyrrolyl, imidazolyl, andtriazolyl, each of which is optionally substituted as described herein(e.g., as in (VII6)).

(VIII1) In one embodiment, L₁ is a bond or a straight or branchedbivalent C₁-C₆ alkylene chain.

(VIII2) In one embodiment, L₁ is —C(O)—, —C(O)(CH₂)_(n)—, —C(O)NH—,—C(O)NH(CH₂)_(n)—, —C(O)N(C₁-C₆ alkyl)-, —C(O)N(C₁-C₆ alkyl)(CH₂)_(n)—,—C(O)O—, —C(O)O(CH₂)_(n)—, —C(S)—, —C(S)(CH₂)_(n)—, —C(S)NH—,—C(S)N(C₁-C₆ alkyl)-, —C(S)NH(CH₂)_(n)—, —C(S)N(C₁-C₆ alkyl)(CH₂)_(n)—,—(CH₂)_(p)NH—, —(CH₂)_(p)N(C₁-C₆ alkyl)-, or —(CH₂)_(p)O—.

(VIII3) In one embodiment, L₁ is a bond.

(VIII4) In one embodiment, L₁ is a straight or branched bivalent C₁-C₆alkylene chain.

(VIII5) In one embodiment, L₁ is —C(O)—.

(VIII6) In one embodiment, L₁ is —C(O)(CH₂)_(n)—.

(VIII7) In one embodiment, L₁ is —C(O)NH—.

(VIII8) In one embodiment, L₁ is —C(O)NH(CH₂)_(n)—.

(VIII9) In one embodiment, L₁ is —C(O)N(C₁-C₆ alkyl)-.

(VIII10) In one embodiment, L₁ is —C(O)N(C₁-C₆ alkyl)(CH₂)_(n)—.

(VIII11) In one embodiment, L₁ is —C(O)O—.

(VIII12) In one embodiment, L₁ is —C(O)O(CH₂)_(n)—.

(VIII13) In one embodiment, L₁ is —C(S)—.

(VIII14) In one embodiment, L₁ is —C(S)(CH₂)_(n)—.

(VIII15) In one embodiment, L₁ is —C(S)NH—.

(VIII16) In one embodiment, L₁ is —C(S)N(C₁-C₆ alkyl)-.

(VIII17) In one embodiment, L₁ is —C(S)NH(CH₂)_(n)—.

(VIII18) In one embodiment, L₁ is —C(S)N(C₁-C₆ alkyl)(CH₂)_(n)—.

(VIII19) In one embodiment, L₁ is —(CH₂)_(p)NH—.

(VIII20) In one embodiment, L₁ is —(CH₂)_(p)N(C₁-C₆ alkyl)-.

(VIII21) In one embodiment, L₁ is —(CH₂)_(p)O—.

(IX1) In one embodiment, n is 1.

(IX2) In one embodiment, n is 2.

(X1) In one embodiment, p is 1 or 2.

(X2) In one embodiment, p is 1.

(X3) In one embodiment, p is 2 or 3.

(X4) In one embodiment, p is 2.

(X5) In one embodiment, p is 3.

(XI1) In one embodiment, X₁, X₂, X3, and X₄ are each CR₄.

(XI2) In one embodiment, one of X₁, X₂, X₃, and X₄ is N, and theremainder of X₁, X₂, X₃, and X₄ are each CR₄.

(XI3) In one embodiment, two of X₁, X₂, X₃, and X₄ are N, and theremainder of X₁, X₂, X₃, and X₄ are each CR₄.

(XI4) In one embodiment, X₁ is N, and X₂, X₃, and X₄ are each CR₄.

(XI5) In one embodiment, X₂ is N, and X₁, X₃, and X₄ are each CR₄.

(XI6) In one embodiment, X₃ is N, and X₁, X₂, and X₄ are each CR₄.

(XI7) In one embodiment, X₄ is N, and X₁, X₂, and X₃ are each CR₄.

(XI8) In one embodiment, X₁ and X₂ are each N, and X₃ and X₄ are eachCR₄.

(X19) In one embodiment, X₁ and X₃ are each N, and X₂ and X₄ are eachCR₄.

(XI10) In one embodiment, X₁ and X₄ are each N, and X₂ and X₃ are eachCR₄.

(XI11) In one embodiment, X₂ and X₃ are each N, and X₁ and X₄ are eachCR₄.

(XI12) In one embodiment, X₂ and X₄ are each N, and X₁ and X₃ are eachCR₄.

(XI13) In one embodiment, X₃ and X₄ are each N, and X₁ and X₂ are eachCR₄.

(XII1) In one embodiment, each R₄ is H.

(XII2) In one embodiment, at least one R₄ is NR₅R₆, NR₇C(O)R₈, C₁-C₆straight-chain or C₃-C₆ branched alkyl (e.g., methyl, ethyl, n-propyl,i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, pentyl, or hexyl), C₁-C₆straight-chain or C₃-C₆ branched haloalkyl (e.g., methyl, ethyl,n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, pentyl, orhexyl, each of which is substituted with one or more halogen (e.g., F,Cl, Br, or I)), C₁-C₆ straight-chain or C₁-C₆ branched hydroxyalkyl(e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl,t-butyl, pentyl, or hexyl, each of which is substituted with one or moreOH), C₁-C₆ straight-chain or C₃-C₆ branched alkoxy (e.g., methoxy,ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy, t-butoxy,pentoxy, or hexyloxy), C₁-C₆ straight-chain or C₃-C₆ branched haloalkoxy(e.g., methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy,s-butoxy, t-butoxy, pentoxy, or hexyloxy, each of which is substitutedwith one or more halogen (e.g., F, Cl, Br, or I)), OH, halogen (e.g., F,Cl, Br, or I), CN, phenyl, C₃-C₆ cycloalkyl, 5- or 6-membered heteroarylcomprising 1-3 heteroatoms selected from N, O, and S, or 5- or6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O,and S, wherein the phenyl, cycloalkyl, heteroaryl, or heterocyclyl isoptionally substituted with one or more substituents independentlyselected from C₁-C₆ straight-chain or C₃-C₆ branched alkyl (e.g.,methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl,pentyl, or hexyl), C₁-C₆ straight-chain or C₃-C₆ branched haloalkyl(e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl,t-butyl, pentyl, or hexyl, each of which is substituted with one or morehalogen (e.g., F, Cl, Br, or I)), C₁-C₆ straight-chain or C₃-C₆ branchedhydroxyalkyl (e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl,s-butyl, t-butyl, pentyl, or hexyl, each of which is substituted withone or more OH), C₁-C₆ straight-chain or C₃-C₆ branched alkoxy (e.g.,methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy,t-butoxy, pentoxy, or hexyloxy), C₁-C₆ straight-chain or C₃-C₆ branchedhaloalkoxy (e.g., methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy,i-butoxy, s-butoxy, t-butoxy, pentoxy, or hexyloxy, each of which issubstituted with one or more halogen (e.g., F, Cl, Br, or I)), OH, andhalogen (e.g., F, Cl, Br, or I).

(XII3) In one embodiment, at least one R₄ is NR₅R₆, NR₇C(O)R₈, C₁-C₆straight-chain or C₃-C₆ branched alkyl (e.g., methyl, ethyl, n-propyl,i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, pentyl, or hexyl), C₁-C₆straight-chain or C₃-C₆ branched haloalkyl (e.g., methyl, ethyl,n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, pentyl, orhexyl, each of which is substituted with one or more halogen (e.g., F,Cl, Br, or I)), C₁-C₆ straight-chain or C₃-C₆ branched hydroxyalkyl(e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl,t-butyl, pentyl, or hexyl, each of which is substituted with one or moreOH), C₁-C₆ straight-chain or C₃-C₆ branched alkoxy (e.g., methoxy,ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy, t-butoxy,pentoxy, or hexyloxy), C₁-C₆ straight-chain or C₃-C₆ branched haloalkoxy(e.g., methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy,s-butoxy, t-butoxy, pentoxy, or hexyloxy, each of which is substitutedwith one or more halogen (e.g., F, Cl, Br, or I)), OH, halogen (e.g., F,Cl, Br, or I), or CN.

(XII4) In one embodiment, one or more R₄ is NR₅R₆.

(XII5) In one embodiment, one or more R₄ is NR₇C(O)R₈

(XII6) In one embodiment, one or more R₄ is C₁-C₆ alkyl, e.g., methyl,ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, pentyl,or hexyl.

(XII7) In one embodiment, one or more R₄ is C₁-C₆ haloalkyl, e.g.,methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl,pentyl, or hexyl, each of which is substituted with one or more halogen(e.g., F, Cl, Br, or I). In one embodiment, one or more R₄ is CF₃, CHF₂,or CH₂F.

(XII8) In one embodiment, one or more R₄ is C₁-C₆ hydroxyalkyl, e.g.,methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl,pentyl, or hexyl, each of which is substituted with one or more OH. Inone embodiment, one or more R₄ is CH₂OH. CH₂CH₂OH, or C(CH₃)₂OH.

(XII9) In one embodiment, one or more R₄ is C₁-C₆ alkoxy, e.g., methoxy,ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy, t-butoxy,pentoxy, or hexyloxy. In one embodiment, at least one R₄ is methoxy.

(XII10) In one embodiment, one or more R₄ is C₁-C₆ haloalkoxy, e.g.,methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy,t-butoxy, pentoxy, or hexyloxy, each of which is substituted with one ormore halogen (e.g., F, Cl, Br, or I). In one embodiment, one or more R₄is OCF₃, OCHF₂, or OCH₂F.

(XII11) In one embodiment, one or more R₄ is OH.

(XII12) In one embodiment, one or more R₄ is halogen. e.g., F, Br, Cl,or I.

(XII13) In one embodiment, one or more R₄ is CN.

(XII14) In one embodiment, one or more R₄ is phenyl, C₃-C₆ cycloalkyl,5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from N,O, and S, or 5- or 6-membered heterocyclyl comprising 1-3 heteroatomsselected from N, O, and S, wherein the phenyl, cycloalkyl, heteroaryl,or heterocyclyl is optionally substituted as described herein (e.g., asin (XII1)).

(XII15) In one embodiment, one or more R₄ is phenyl, optionallysubstituted as described herein (e.g., as in (XII1)).

(XII16) In one embodiment, one or more R₄ is C₃-C₆ cycloalkyl,optionally substituted as described herein (e.g., as in (XII1)). In oneembodiment, one or more R₄ is cyclopropyl. In one embodiment, one ormore R₄ is cyclobutyl. In one embodiment, one or more R₄ is cyclopentyl.In one embodiment, one or more R₄ is cyclohexyl.

(XII17) In one embodiment, one or more R₄ is 5- or 6-membered heteroarylcomprising 1-3 heteroatoms selected from N, O, and S, optionallysubstituted as described herein (e.g., as in (XIII)).

(XII18) In one embodiment, one or more R₄ is 5-membered heteroarylcomprising 1-3 heteroatoms selected from N, O, and S, optionallysubstituted as described herein (e.g., as in (XII1)). In one embodiment,one or more R₄ is 5-membered heteroaryl comprising 1-2 heteroatomsselected from N, O, and S. In one embodiment, one or more R₄ is5-membered heteroaryl comprising 1-2 heteroatoms selected from N and O.In one embodiment, one or more R₄ is heteroaryl selected from pyrrolyl,furanyl, thiophenyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl,thiazolyl, isothiazolyl, triazolyl, oxadiazolyl, and thiadiazolyl, eachof which is optionally substituted as described herein (e.g., as in(XII1)). In one embodiment, one or more R₄ is pyrazolyl or imidazolyl,optionally substituted as described herein (e.g., as in (XII1)).

(XII19) In one embodiment, one or more R₄ is 6-membered heteroarylcomprising 1-3 heteroatoms selected from N, O, and S, optionallysubstituted as described herein (e.g., as in (XII1)). In one embodiment,one or more R, is 6-membered heteroaryl comprising 1-2 heteroatomsselected from N, O, and S. In one embodiment, one or more R₄ is6-membered heteroaryl comprising 1-2 heteroatoms selected from N and O.In one embodiment, one or more R₄ is heteroaryl selected from pyridinyl,pyridazinyl, pyrimidinyl, pyrazinyl, pyranyl, thiopyranyl, diazinyl,thiazinyl, dioxinyl, and triazinyl, each of which is optionallysubstituted as described herein (e.g., as in (XII1)). In one embodiment,one or more R₄ is 6-membered heteroaryl comprising 1-2 nitrogen atoms.In one embodiment, one or more R₄ is 6-membered heteroaryl comprising 1nitrogen atom. In one embodiment, one or more R₄ is 6-memberedheteroaryl comprising 2 nitrogen atoms. In one embodiment, one or moreR₄ is heteroaryl selected from pyridinyl, pyridazinyl, and pyrimidinyl,each of which is optionally substituted as described herein (e.g., as in(XII1)).

(XII20) In one embodiment, one or more R₄ is 5- or 6-memberedheterocyclyl comprising 1-3 heteroatoms selected from N, O, and S,optionally substituted as described herein (e.g., as in (XII1)).

(XII21) In one embodiment, one or more R₄ is 5-membered heterocyclylcomprising 1-3 heteroatoms selected from N, O, and S, optionallysubstituted as described herein (e.g., as in (XII1)). In one embodiment,one or more R₄ is 5-membered heterocyclyl comprising 1-2 heteroatomsselected from N, O, and S. In one embodiment, one or more R₄ is5-membered heterocyclyl comprising 1-2 heteroatoms selected from N andO. In one embodiment, one or more R₄ is 5-membered heterocyclylcomprising 1 heteroatom selected from N, O, and S. In one embodiment,one or more R₄ is heterocyclyl selected from pyrrolidinyl,tetrahydrofuranyl, tetrahydrothiophenyl, pyrazolidinyl, imidazolidinyl,oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolidinyl,triazolidinyl, oxadiazolidinyl, isoxadiazolidinyl, thiadiazolidinyl, andisothiadiazolidinyl, each of which is optionally substituted asdescribed herein (e.g., as in (XII1))

(XII22) In one embodiment, one or more R₄ is 6-membered heterocyclylcomprising 1-3 heteroatoms selected from N, O, and S, optionallysubstituted as described herein (e.g., as in (XII1)). In one embodiment,one or more R₄ is 6-membered heterocyclyl comprising 1-2 heteroatomsselected from N, O, and S. In one embodiment, one or more R₄ is6-membered heterocyclyl comprising 1-2 heteroatoms selected from N andO. In one embodiment, one or more R₄ is 6-membered heterocyclylcomprising 1 heteroatom selected from N, O, and S. In one embodiment,one or more R₄ is heterocyclyl selected from piperidinyl, piperazinyl,tetrahydropyranyl, hexahydropyridazinyl, hexahydropyrimidinyl,morpholinyl, and triazinanyl, each of which is optionally substituted asdescribed herein (e.g., as in (XII1)). In one embodiment, one or more R;is piperidinyl or piperazinyl, each of which is optionally substitutedas described herein (e.g., as in (XII1)).

(XII11) In one embodiment, each R₅ and each R₆ are H.

(XII12) In one embodiment, at least one R₅ and at least one R₆ are C₁-C₄straight-chain or C₃-C₄ branched alkyl (e.g., methyl, ethyl, n-propyl,i-propyl, n-butyl, i-butyl, s-butyl, or t-butyl).

(XIII3) In one embodiment, each R₅ is H and at least one R₆ is C₁-C₄straight-chain or C₃-C₄ branched alkyl (e.g., methyl, ethyl, n-propyl,i-propyl, n-butyl, i-butyl, s-butyl, or t-butyl).

(XIII4) In one embodiment, at least one R₅ is C₁-C₄ straight-chain orC₃-C₄ branched alkyl (e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl,i-butyl, s-butyl, ort-butyl) and each R₆ is H.

(XIII5) In one embodiment, one or more R₄ is NR₅R₆, wherein each of R₅and R₆ is H.

(XIII6) In one embodiment, one or more R₄ is NR₅R₆, wherein each of R₅and R₆ is C₁-C₄ alkyl. In one embodiment, R₄ is N(CH₃)₂.

(XIII7) In one embodiment, one or more R₄ is NR₅R₆, wherein R₅ is H andR₆ is C₁-C₄ alkyl. In one embodiment, R₄ is NHCH₃.

(XIV1) In one embodiment, each R₇ is H.

(XIV2) In one embodiment, at least one R₇ is C₁-C₄ straight-chain orC₃-C₄ branched alkyl (e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl,i-butyl, s-butyl, or t-butyl).

(XV1) In one embodiment, each R₈ is independently C₁-C₄ straight-chainor C₃-C₄ branched alkyl (e.g., methyl, ethyl, n-propyl, i-propyl,n-butyl, i-butyl, s-butyl, or t-butyl).

(XV2) In one embodiment, one or more R₄ is NR₇C(O)R₈, wherein R₇ is Hand R₈ is C₁-C₄ alkyl.

(XV3) In one embodiment, one or more R₄ is NR₇C(O)R₈, wherein each of R₇and R₈ is C₁-C₄ alkyl.

(XVI1) In one embodiment, none of X₅, X₆, X₇, and X₈ is N, one of X₅,X₆, X₇, and X₈ is CR₁₀, and three of X₅, X₆, X₇, and X₈ are eachindependently CR₉.

(XVI2) In one embodiment, none of X₅, X₆, X₇, and X₈ is N, two of X₅,X₆, X₇, and X₈ are each independently CR₁₀, and two of X₅, X₆, X₇, andX₈ are each independently CR₉.

(XVI3) In one embodiment, none of X₅, X₆, X₇, and X₈ is N, three of X₅,X₆, X₇, and X₈ are each independently CR₁₀, and one of X₅, X₆, X₇, andX₈ is CR₉.

(XVI4) In one embodiment, one of X₅, X₆, X₇, and X₈ is N, one of X₅, X₆,X₇, and X₈ is CR₁₀, and two of X₅, X₆, X₇, and X₈ are each independentlyCR₉.

(XVI5) In one embodiment, one of X₅, X₆, X₇, and X₈ is N, two of X₅, X₆,X₇, and X₈ are each independently CR₁₀, and one of X₅, X₆, X₇, and X₈ isCR₉.

(XVI6) In one embodiment, two of X₅, X₆, X₇, and X₈ is N, one of X₅, X₆,X₇, and X₈ is CR₁₀, and one of X₅, X₆, X₇, and X₈ is CR₉.

(XVI7) In one embodiment, X₆, X₇, and X₈ are CR₉, and X₅ is CR₁₀.

(XVI8) In one embodiment, X₅, X₇, and X₈ are CR₉, and X₆ is CR₁₀.

(XVI9) In one embodiment, X₅, X₆, and X₈ are CR₉, and X₇ is CR₁₀.

(XVI10) In one embodiment, X₅, X₆, and X₇ are CR₉, and X₈ is CR₁₀.

(XVI11) In one embodiment, X₇ and X₈ are CR₉, and X₅ and X₆ are CR₁₀.

(XVI12) In one embodiment, X₆ and X₈ are CR₉, and X₅ and X₇ are CR₁₀.

(XVI13) In one embodiment, X₅ and X₈ are CR₉, and X₆ and X₇ are CR₁₀.

(XVI14) In one embodiment, X₆ and X₇ are CR₉, and X₅ and X₈ are CR₁₀.

(XVI15) In one embodiment, X₅ and X₇ are CR₉, and X₆ and X₈ are CR₁₀.

(XVI16) In one embodiment, X₅ and X₆ are CR₉, and X₇ and X₈ are CR₁₀.

(XVI17) In one embodiment, X₆, X₇, and X₈ are CR₁₀, and X₅ is CR₉.

(XVI18) In one embodiment, X₅, X₇, and X₈ are CR₁₀, and X₆ is CR₉.

(XVI19) In one embodiment, X₅, X₆, and X₈ are CR₁₀, and X₇ is CR₉.

(XVI20) In one embodiment, X₅, X₆, and X₇ are CR₁₀, and X₈ is CR₉.

(XVI21) In one embodiment, X₈ is N, X₆ and X, are CR₉, and X₅ is CR₁₀.

(XVI22) In one embodiment, X₈ is N, X₅ and X₇ are CR₉, and X₆ is CR₁₀.

(XVI23) In one embodiment, X₈ is N, X₅ and X₆ are CR₉, and X₇ is CR₁₀.

(XVI24) In one embodiment, X₇ is N, X₆ and X₈ are CR₉, and X₅ is CR₁₀.

(XVI25) In one embodiment, X₇ is N, X₅ and X₈ are CR₉, and X₆ is CR₁₀.

(XVI26) In one embodiment, X₇ is N, X₅ and X₆ are CR₉, and X₈ is CR₁₀.

(XVI27) In one embodiment, X₆ is N, X₇ and X₈ are CR₉, and X₅ is CR₁₀.

(XVI28) In one embodiment, X₆ is N, X₅ and X₈ are CR₉, and X₇ is CR₁₀.

(XVI29) In one embodiment, X₆ is N, X₅ and X₇ are CR₉, and X₈ is CR₁₀.

(XVI30) In one embodiment, X₅ is N, X₇ and X₈ are CR₉, and X₆ is CR₁₀.

(XVI31) In one embodiment, X₅ is N, X₆ and X₈ are CR₉, and X₇ is CR₁₀.

(XVI32) In one embodiment, X₅ is N, X₆ and X₇ are CR₉, and X₈ is CR₁₀.

(XVI33) In one embodiment, X₈ is N, X₆ and X₇ are CR₁₀, and X₅ is CR₉.

(XVI34) In one embodiment, X₈ is N, X₅ and X₇ are CR₁₀, and X₆ is CR₉.

(XVI35) In one embodiment, X₈ is N, X₅ and X₆ are CR₁₀, and X₇ is CR₉.

(XVI36) In one embodiment, X₇ is N, X₆ and X₈ are CR₁₀, and X₅ is CR₉.

(XVI37) In one embodiment, X₇ is N, X₅ and X₈ are CR₁₀, and X₆ is CR₉.

(XVI38) In one embodiment, X₇ is N, X₅ and X₆ are CR₁₀, and X₈ is CR₉.

(XVI39) In one embodiment, X₆ is N, X₇ and X₅ are CR₁₀, and X₆ is CR₉.

(XVI40) In one embodiment, X₆ is N, X₅ and X₅ are CR₁₀, and X₇ is CR₉.

(XVI41) In one embodiment, X₆ is N, X₅ and X₇ are CR₁₀, and X₈ is CR₉.

(XVI42) In one embodiment, X₅ is N, X₇ and X₈ are CR₁₀, and X₆ is CR₉.

(XVI43) In one embodiment, X₅ is N, X₆ and X₈ are CR₁₀, and X₇ is CR₉.

(XVI44) In one embodiment, X₅ is N, X₆ and X₇ are CR₁₀, and X₈ is CR₉.

(XVI45) In one embodiment, X₈ and X₅ are N, X₆ is CR₉, and X₇ is CR₁₀.

(XVI46) In one embodiment, X₈ and X₅ are N, X₇ is CR₉, and X₆ is CR₁₀.

(XVI47) In one embodiment, X₈ and X₆ are N, X₇ is CR₉, and X₅ is CR₁₀.

(XVI48) In one embodiment, X₈ and X₆ are N, X₅ is CR₉, and X₇ is CR₁₀.

(XVI49) In one embodiment, X₈ and X₇ are N, X₅ is CR₉, and X₆ is CR₁₀.

(XVI50) In one embodiment, X₈ and X₇ are N, X₆ is CR₉, and X₅ is CR₁₀.

(XVI51) In one embodiment, X₇ and X₆ are N, X₅ is CR₉, and X₈ is CR₁₀.

(XVI52) In one embodiment, X₇ and X₆ are N, X₈ is CR₉, and X₅ is CR₁₀.

(XVI53) In one embodiment, X₇ and X₅ are N, X₆ is CR₉, and X₈ is CR₁₀.

(XVI54) In one embodiment, X₇ and X₅ are N, X₈ is CR₉, and X₆ is CR₁₀.

(XVI55) In one embodiment, X₆ and X₅ are N, X₈ is CR₉, and X₇ is CR₁₀.

(XVI56) In one embodiment, X₆ and X₅ are N, X₇ is CR₉, and X₈ is CR₁₀.

(XVII1) In one embodiment, each R₉ is H.

(XVII2) In one embodiment, at least one R₉ is C₁-C₆ straight-chain orC₃-C₆ branched alkyl (e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl,i-butyl, s-butyl, t-butyl, pentyl, or hexyl), C₁-C₆ straight-chain orC₃-C₆ branched haloalkyl (e.g., methyl, ethyl, n-propyl, i-propyl,n-butyl, i-butyl, s-butyl, t-butyl, pentyl, or hexyl, each of which issubstituted with one or more halogen (e.g., F, Cl, Br, or I)), C₁-C₆straight-chain or C₃-C₆ branched hydroxyalkyl (e.g., methyl, ethyl,n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, pentyl, orhexyl, each of which is substituted with one or more OH), C₁-C₆straight-chain or C₃-C₆ branched alkoxy (e.g., methoxy, ethoxy,n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy, t-butoxy, pentoxy,or hexyloxy), C₁-C₆ straight-chain or C₃-C₆ branched haloalkoxy (e.g.,methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy,t-butoxy, pentoxy, or hexyloxy, each of which is substituted with one ormore halogen (e.g., F, Cl, Br, or I)), OH, halogen (e.g., F, Cl, Br, orI), or CN.

(XVII3) In one embodiment, one or more R₉ is C₁-C₆ alkyl, e.g., methyl,ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, pentyl,or hexyl.

(XVII4) In one embodiment, one or more R₉ is C₁-C₆ haloalkyl, e.g.,methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl,pentyl, or hexyl, each of which is substituted with one or more halogen(e.g., F, Cl, Br, or I). In one embodiment, one or more R₉ is CF₃, CHF₂,or CH₂F.

(XVII5) In one embodiment, one or more R₉ is C₁-C₆ hydroxyalkyl, e.g.,methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl,pentyl, or hexyl, each of which is substituted with one or more OH. Inone embodiment, one or more R₉ is CH₂OH, CH₂CH₂OH, or C(CH₃)₂OH.

(XVII6) In one embodiment, one or more R₉ is C₁-C₆ alkoxy, e.g.,methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy,t-butoxy, pentoxy, or hexyloxy. In one embodiment, at least one R₉ ismethoxy.

(XVII7) In one embodiment, one or more R₉ is C₁-C₆ haloalkoxy, e.g.,methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy,t-butoxy, pentoxy, or hexyloxy, each of which is substituted with one ormore halogen (e.g., F, Cl, Br, or I). In one embodiment, one or more R₉is OCF₃, OCHF₂, or OCH₂F.

(XVII8) In one embodiment, one or more R₉ is OH.

(XVII9) In one embodiment, one or more R₉ is halogen, e.g., F, Br, Cl,or I.

(XVII10) In one embodiment, one or more R₉ is CN.

(XII1) In one embodiment, each R₁₀ is independently phenyl, C₃-C₆cycloalkyl, 5- or 6-membered heteroaryl comprising 1-4 heteroatomsselected from N, O, and S, or 5- or 6-membered heterocyclyl comprising1-3 heteroatoms selected from N, O, and S, wherein the phenyl,cycloalkyl, heteroaryl, or heterocyclyl is optionally substituted withone or more R₁₁.

(XIII2) In one embodiment, at least one R₁₀ is phenyl, optionallysubstituted with one or more R₁₁.

(XIII3) In one embodiment, at least one R₁₀ is C₁-C₆ cycloalkyl,optionally substituted with one or more R₁₁. In one embodiment, at leastone R₁₀ is cyclopropyl. In one embodiment, at least one R₁₀ iscyclobutyl. In one embodiment, at least one R₁₀ is cyclopentyl. In oneembodiment at least one R₁₀ is cyclohexyl.

(XIII4) In one embodiment, at least one R₁₀ is 5- or 6-memberedheteroaryl comprising 1-4 heteroatoms selected from N, O, and S,optionally substituted with one or more R₁₁. In one embodiment, at leastone R₁₀ is 5- or 6-membered heteroaryl comprising 1-3 heteroatomsselected from N, O, and S. In one embodiment, at least one R₁₀ is 5- or6-membered heteroaryl comprising 1-2 heteroatoms selected from N, O, andS. In one embodiment, at least one R₁₀ is 5- or 6-membered heteroarylcomprising 1 heteroatom selected from N, O, and S.

(XIII5) In one embodiment, at least one R₁₀ is 5-membered heteroarylcomprising 1-4 heteroatoms selected from N, O, and S, optionallysubstituted with one or more R₁₁. In one embodiment at least one R₁₀ is5-membered heteroaryl comprising 1-3 heteroatoms selected from N, O, andS. In one embodiment at least one R₁₀ is 5-membered heteroarylcomprising 1-2 heteroatoms selected from N, O, and S. In one embodiment,at least one R₁₀ is 5-membered heteroaryl comprising 1 heteroatomselected from N, O, and S. In one embodiment, at least one R₁₀ isheteroaryl selected from pyrrolyl, furanyl, thiophenyl, pyrazolyl,imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, triazolyl,oxadiazolyl, thiadiazolyl, and tetrazolyl, each of which is optionallysubstituted with one or more R₁₁. In one embodiment, at least one R₁₀ istetrazolyl optionally substituted with one or more R₁₁.

(XIII6) In one embodiment, at least one R₁₀ is 6-membered heteroarylcomprising 1-4 heteroatoms selected from N, O, and S, optionallysubstituted with one or more R₁₁. In one embodiment at least one R₁₀ is6-membered heteroaryl comprising 1-3 heteroatoms selected from N, O, andS. In one embodiment at least one R₁₀ is 6-membered heteroarylcomprising 1-2 heteroatoms selected from N, O, and S. In one embodiment,at least one R₁₀ is 6-membered heteroaryl comprising 1 heteroatomselected from N, O, and S. In one embodiment, at least one R₁₀ isheteroaryl selected from pyridinyl, pyridazinyl, and pyrimidinyl, eachof which is optionally substituted with one or more R₁₁.

(XIII7) In one embodiment, at least one R₁₀ is 5- or 6-memberedheterocyclyl comprising 1-3 heteroatoms selected from N, O, and S,optionally substituted with one or more R₁₁. In one embodiment, at leastone R₁₀ is 5- or 6-membered heterocyclyl comprising 1-2 heteroatomsselected from N, O, and S. In one embodiment, at least one R₁₀ is 5- or6-membered heterocyclyl comprising 1 heteroatom selected from N, O, andS.

(XIII8) In one embodiment, at least one R₁₀ is 5-membered heterocyclylcomprising 1-3 heteroatoms selected from N, O, and S, optionallysubstituted with one or more R₁₁. In one embodiment, at least one R₁₀ is5-membered heterocyclyl comprising 1-2 heteroatoms selected from N, O,and S. In one embodiment, at least one R₁₀ is 5-membered heterocyclylcomprising 1 heteroatom selected from N, O, and S. In one embodiment, atleast one R₁₀ is heterocyclyl selected from dihydropyrrolyl,pyrrolidinyl, dihydroimidazolyl, imidazolinyl, dihydrooxazolyl,oxazolidinyl, dihydroisoxazolyl, isoxazolidinyl, dihydrothiazolyl,thiazolidinyl, dihydroisothiazolyl, isothiazolidinyl, dihydrofuryl,tetrahydrofuranyl, dioxolanyl, dihydrothiophenyl, tetrahydrothiophenyl,and dithiolanyl, each of which is optionally substituted with one ormore R₁₁.

(XIII9) In one embodiment, at least one R₁₀ is 6-membered heterocyclylcomprising 1-3 heteroatoms selected from N, O, and S, optionallysubstituted with one or more R₁₁. In one embodiment, at least one R₁₀ is6-membered heterocyclyl comprising 1-2 heteroatoms selected from N, O,and S. In one embodiment, at least one R₁₀ is 6-membered heterocyclylcomprising 1 heteroatom selected from N, O, and S. In one embodiment, atleast one R₁₀ is dihydropyridinyl, tetrahydropyridinyl, piperidinyl,dihydropyrazinyl, tetrahydropyrazinyl, piperazinyl, morpholinyl,oxazinyl, dihydrooxazinyl, oxazinanyl, dihydrothiazinyl,tetrahydrothiazinyl, thiomorpholinyl, pyranyl, dihydropyranyl,tetrahydropyranyl, dioxinyl, dihydrodioxinyl, dioxanyl, thiopyranyl,dihydrothiopyranyl, tetrahydropyranyl, and dithianyl, each of which isoptionally substituted with one or more R₁₁.

(XIII10) In one embodiment, at least one R₁₀ is piperazinyl.

(XIII11) In one embodiment, at least one R₁₀ is piperazinyl substitutedwith methyl.

(XIII12) In one embodiment, at least one R₁₀ is pyrazolyl.

(XIII13) In one embodiment, at least one R₁₀ is pyrazolyl substitutedwith methyl.

(XIII14) In one embodiment, at least one R₁₀ is morpholinyl.

(XIII15) In one embodiment, at least one R₁₀ is tetrahydropyridinyl. Inone embodiment, R₁₀ is 1,2,3,6-tetrahydropyridinyl.

(XIII16) In one embodiment, at least one R₁₀ is tetrazolyl.

(XIV1) In one embodiment, each R₁₁ is independently C₁-C₆ straight-chainor C₃-C₆ branched alkyl (e.g., methyl, ethyl, n-propyl, i-propyl,n-butyl, i-butyl, s-butyl, t-butyl, pentyl, or hexyl), C₁-C₆straight-chain or C₃-C₆ branched haloalkyl (e.g., methyl, ethyl,n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, pentyl, orhexyl, each of which is substituted with one or more halogen (e.g., F,Cl, Br, or I)), C₁-C₆ straight-chain or C₃-C₆ branched hydroxyalkyl(e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl,t-butyl, pentyl, or hexyl, each of which is substituted with one or moreOH), C₁-C₆ straight-chain or C₃-C₆ branched alkoxy (e.g., methoxy,ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy, t-butoxy,pentoxy, or hexyloxy), C₁-C₆ straight-chain or C₃-C₆ branched haloalkoxy(e.g., methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy,s-butoxy, t-butoxy, pentoxy, or hexyloxy, each of which is substitutedwith one or more halogen (e.g., F, Cl, Br, or I)). OH, halogen (e.g., F,Cl, Br, or I), CN, phenyl, C₃-C₆ cycloalkyl, 5- or 6-membered heteroarylcomprising 1-3 heteroatoms selected from N, O, and S, or 5- or6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O,and S, wherein the phenyl, cycloalkyl, heteroaryl, or heterocyclyl isoptionally substituted with one or more substituents independentlyselected from C₁-C₆ straight-chain or C₃-C₆ branched alkyl (e.g.,methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl,pentyl, or hexyl), C₁-C₆ straight-chain or C₃-C₆ branched haloalkyl(e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl,t-butyl, pentyl, or hexyl, each of which is substituted with one or morehalogen (e.g., F, Cl, Br, or I)), C₁-C₆ straight-chain or C₃-C₆ branchedhydroxyalkyl (e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl,s-butyl, t-butyl, pentyl, or hexyl, each of which is substituted withone or more OH), C₁-C₆ straight-chain or C₃-C₆ branched alkoxy (e.g.,methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy,t-butoxy, pentoxy, or hexyloxy), C₁-C₆ straight-chain or C₃-C₆ branchedhaloalkoxy (e.g., methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy,i-butoxy, s-butoxy, t-butoxy, pentoxy, or hexyloxy, each of which issubstituted with one or more halogen (e.g., F, Cl, Br, or I)), OH, andhalogen (e.g., F, Cl, Br, or I).

(XIV2) In one embodiment, each R₁₁ is independently C₁-C₆ straight-chainor C₃-C₆ branched alkyl (e.g., methyl, ethyl, n-propyl, i-propyl,n-butyl, i-butyl, s-butyl, t-butyl, pentyl, or hexyl), C₁-C₆straight-chain or C₃-C₆ branched haloalkyl (e.g., methyl, ethyl,n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, pentyl, orhexyl, each of which is substituted with one or more halogen (e.g., F,Cl, Br, or I)), C₁-C₆ straight-chain or C₃-C₆ branched hydroxyalkyl(e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl,t-butyl, pentyl, or hexyl, each of which is substituted with one or moreOH), C₁-C₆ straight-chain or C₃-C₆ branched alkoxy (e.g., methoxy,ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy, t-butoxy,pentoxy, or hexyloxy), C₁-C₆ straight-chain or C₃-C₆ branched haloalkoxy(e.g., methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy,s-butoxy, t-butoxy, pentoxy, or hexyloxy, each of which is substitutedwith one or more halogen (e.g., F, Cl, Br, or I)), OH, halogen (e.g., F,Cl, Br, or I), or CN.

(XIV3) In one embodiment, one or more R₁₁ is C₁-C₆ alkyl, e.g., methyl,ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, pentyl,or hexyl.

(XV4) In one embodiment, one or more R₁₁ is C₁-C₆ haloalkyl, e.g.,methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl,pentyl, or hexyl, each of which is substituted with one or more halogen(e.g., F, Cl, Br, or I). In one embodiment, one or more R₁₁ is CF₃,CHF₂, or CH₂F.

(XIV5) In one embodiment, one or more R₁₁ is C₁-C₆ hydroxyalkyl, e.g.,methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl,pentyl, or hexyl, each of which is substituted with one or more OH. Inone embodiment, one or more R₁₁ is CH₂OH. CH₃CH₂OH, or C(CH₃)₂OH.

(XIV6) In one embodiment, one or more R₁₁ is C₁-C₆ alkoxy, e.g.,methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy,t-butoxy, pentoxy, or hexyloxy. In one embodiment, at least one R₁₁ ismethoxy.

(XIV7) In one embodiment, one or more R₀ is C₁-C₆ haloalkoxy, e.g.,methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy,t-butoxy, pentoxy, or hexyloxy, each of which is substituted with one ormore halogen (e.g., F, Cl, Br, or I). In one embodiment, one or more R₁₁is OCF₃, OCHF₂, or OCH₂F.

(XIV8) In one embodiment, one or more R₁₁ is OH.

(XIV9) In one embodiment, one or more R₁₁ is halogen, e.g., F, Br, Cl,or I.

(XIV10) In one embodiment, one or more R₁₁ is CN.

(XIV11) In one embodiment, one or more R₁₁ is phenyl, C₃-C₆ cycloalkyl,5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from N,O, and S, or 5- or 6-membered heterocyclyl comprising 1-3 heteroatomsselected from N, O, and S, wherein the phenyl, cycloalkyl, heteroaryl,or heterocyclyl is optionally substituted as described herein (e.g., asin (XIV1)).

(XIV12) In one embodiment, one or more R₁₁ is phenyl, optionallysubstituted as described herein (e.g., as in (XIV1)).

(XIV13) In one embodiment, one or more R₁₁ is C₁-C₆ cycloalkyl,optionally substituted as described herein (e.g., as in (XIV1)). In oneembodiment, one or more R₁₁ is cyclopropyl. In one embodiment, one ormore R₁₁ is cyclobutyl. In one embodiment, one or more R₁₁ iscyclopentyl. In one embodiment, one or more R₁₁ is cyclohexyl.

(XIV14) In one embodiment, one or more R₁₁ is 5- or 6-memberedheteroaryl comprising 1-3 heteroatoms selected from N, O, and S,optionally substituted as described herein (e.g., as in (XIV1)).

(XIV15) In one embodiment, one or more R₁₁ is 5-membered heteroarylcomprising 1-3 heteroatoms selected from N, O, and S, optionallysubstituted as described herein (e.g., as in (XIV1)). In one embodiment,one or more R₁₁ is 5-membered heteroaryl comprising 1-2 heteroatomsselected from N, O, and S. In one embodiment, one or more R₁₁ is5-membered heteroaryl comprising 1-2 heteroatoms selected from N and O.In one embodiment, one or more R₀ is heteroaryl selected from pyrrolyl,furanyl, thiophenyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl,thiazolyl, isothiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, andtetrazolyl, each of which is optionally substituted as described herein(e.g., as in (XIV1)). In one embodiment, one or more R₁₁ is pyrazolyl orimidazolyl, optionally substituted as described herein (e.g., as in(XIV1)).

(XIV16) In one embodiment, one or more R₁₁ is 6-membered heteroarylcomprising 1-3 heteroatoms selected from N, O, and S, optionallysubstituted as described herein (e.g., as in (XIV1)). In one embodiment,one or more R₁₁ is 6-membered heteroaryl comprising 1-2 heteroatomsselected from N, O, and S. In one embodiment, one or more R₁₁ is6-membered heteroaryl comprising 1-2 heteroatoms selected from N and O.In one embodiment, one or more R₁₁ is heteroaryl selected frompyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, pyranyl, thiopyranyl,diazinyl, thiazinyl, dioxinyl, and triazinyl, each of which isoptionally substituted as described herein (e.g., as in (XIV1)). In oneembodiment, one or more R₁₁ is 6-membered heteroaryl comprising 1-2nitrogen atoms. In one embodiment, one or more R₁₁ is 6-memberedheteroaryl comprising 1 nitrogen atom. In one embodiment, one or moreR₁₁ is 6-membered heteroaryl comprising 2 nitrogen atoms. In oneembodiment, one or more R₁₁ is heteroaryl selected from pyridinyl,pyridazinyl, and pyrimidinyl, each of which is optionally substituted asdescribed herein (e.g., as in (XIV1)).

(XIV17) In one embodiment, one or more R₁₁ is 5- or 6-memberedheterocyclyl comprising 1-3 heteroatoms selected from N, O, and S,optionally substituted as described herein (e.g., as in (XIV1)).

(XIV18) In one embodiment, one or more R₁₁ is 5-membered heterocyclylcomprising 1-3 heteroatoms selected from N, O, and S, optionallysubstituted as described herein (e.g., as in (XIV1)). In one embodiment,one or more R₁₁ is 5-membered heterocyclyl comprising 1-2 heteroatomsselected from N, O, and S. In one embodiment, one or more R₁₁ is5-membered heterocyclyl comprising 1-2 heteroatoms selected from N andO. In one embodiment, one or more R₁₁ is 5-membered heterocyclylcomprising 1 heteroatom selected from N, O, and S. In one embodiment,one or more R₁₁ is heterocyclyl selected from pyrrolidinyl,tetrahydrofuranyl, tetrahydrothiophenyl, pyrazolidinyl, imidazolidinyl,oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolidinyl,triazolidinyl, oxadiazolidinyl, isoxadiazolidinyl, thiadiazolidinyl, andisothiadiazolidinyl, each of which is optionally substituted asdescribed herein (e.g., as in (XIV1)).

(XIV19) In one embodiment, one or more R₁₁ is 6-membered heterocyclylcomprising 1-3 heteroatoms selected from N, O, and S, optionallysubstituted as described herein (e.g., as in (XIV1)). In one embodiment,one or more R₁₁ is 6-membered heterocyclyl comprising 1-2 heteroatomsselected from N, O, and S. In one embodiment, one or more R₁₁ is6-membered heterocyclyl comprising 1-2 heteroatoms selected from N andO. In one embodiment, one or more R₁₁ is 6-membered heterocyclylcomprising 1 heteroatom selected from N, O, and S. In one embodiment,one or more R₁₁ is heterocyclyl selected from piperidinyl, piperazinyl,tetrahydropyranyl, hexahydropyridazinyl, hexahydropyrimidinyl,morpholinyl, and triazinanyl, each of which is optionally substituted asdescribed herein (e.g., as in (XIV1)). In one embodiment, one or moreR₁₁ is piperidinyl or piperazinyl, each of which is optionallysubstituted as described herein (e.g., as in (XIV1)).

(XIV20) In one embodiment, at least one R₁₁ is piperazinyl.

(XIV21) In one embodiment, at least one R₁₁ is piperazinyl substitutedwith methyl.

(XIV22) In one embodiment, at least one R₁₁ is pyrazolyl.

(XIV23) In one embodiment, at least one R₁₁ is pyrazolyl substitutedwith methyl.

(XIV24) In one embodiment, at least one R₁₁ is tetrazolyl.

(XIV25) In one embodiment, at least one R₁₁ is morpholinyl.

(XIV26) In one embodiment, at least one R₁₁ is tetrahydropyridinyl. Inone embodiment, at least one R₁₁ is 1,2,3,6-tetrahydropyridinyl.

(XIV27) In one embodiment, two R₁₁, together with the adjacent atoms towhich they are attached, form phenyl, C₅-C₆ cycloalkyl, 5- or 6-memberedheteroaryl comprising 1-3 heteroatoms selected from N, O, and S, or 5-or 6-membered heterocyclyl comprising 1-3 heteroatoms selected from N,O, and S, wherein the phenyl, cycloalkyl, heteroaryl, or heterocyclyl isoptionally substituted with one or more substituents independentlyselected from C₁-C₆ straight-chain or C₃-C₆ branched alkyl (e.g.,methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl,pentyl, or hexyl), C₁-C₆ straight-chain or C₃-C₆ branched haloalkyl(e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl,t-butyl, pentyl, or hexyl, each of which is substituted with one or morehalogen (e.g., F, Cl, Br, or I)). C₁-C₆ straight-chain or C₃-C₆ branchedhydroxyalkyl (e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl,s-butyl, t-butyl, pentyl, or hexyl, each of which is substituted withone or more OH), C₁-C₆ straight-chain or C₃-C₆ branched alkoxy (e.g.,methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy,t-butoxy, pentoxy, or hexyloxy), C₁-C₆ straight-chain or C₃-C₆ branchedhaloalkoxy (e.g., methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy,i-butoxy, s-butoxy, t-butoxy, pentoxy, or hexyloxy, each of which issubstituted with one or more halogen (e.g., F, Cl, Br, or I)), OH, andhalogen (e.g., F, Cl, Br, or I).

(XIV28) In one embodiment, two R₁₁, taken together with the adjacentatoms to which they are attached, form phenyl, optionally substituted asdescribed herein (e.g., as in (XIV27)).

(XIV29) In one embodiment, two R₁₁, taken together with the adjacentatoms to which they are attached, form C₅-C₆ cycloalkyl (e.g.,cyclopentyl or cyclohexyl), optionally substituted as described herein(e.g., as in (XIV27)).

(XIV30) In one embodiment, two R₁₁, taken together with the adjacentatoms to which they are attached, form 5- or 6-membered heteroarylcomprising 1-3 heteroatoms selected from N, O, and S, optionallysubstituted as described herein (e.g., as in (XIV27)).

(XIV31) In one embodiment, two R₁₁, taken together with the adjacentatoms to which they are attached, form 5-membered heteroaryl comprising1-3 heteroatoms selected from N, O, and S, optionally substituted asdescribed herein (e.g., as in (XIV27)). In one embodiment, two R₁₁,taken together with the adjacent atoms to which they are attached, form5-membered heteroaryl comprising 1-2 heteroatoms selected from N, O, andS. In one embodiment, two R₁₁, taken together with the adjacent atoms towhich they are attached, form 5-membered heteroaryl comprising 1-2heteroatoms selected from N and O. In one embodiment, two R₁₁, takentogether with the adjacent atoms to which they are attached, formheteroaryl selected from pyrrolyl, furanyl, thiophenyl, pyrazolyl,imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, triazolyl,oxadiazolyl, thiadiazolyl, and tetrazolyl, each of which is optionallysubstituted as described herein (e.g., as in (XIV27)). In oneembodiment, two R₁₁, taken together with the adjacent atoms to whichthey are attached, form pyrazolyl or imidazolyl, optionally substitutedas described herein (e.g., as in (XIV27)).

(XIV32) In one embodiment, two R₁₁, taken together with the adjacentatoms to which they are attached, form 6-membered heteroaryl comprising1-3 heteroatoms selected from N, O, and S, optionally substituted asdescribed herein (e.g., as in (XIV27)). In one embodiment, two R₁₁,taken together with the adjacent atoms to which they are attached, form6-membered heteroaryl comprising 1-2 heteroatoms selected from N, O, andS. In one embodiment, two R₁₁, taken together with the adjacent atoms towhich they are attached, form 6-membered heteroaryl comprising 1-2heteroatoms selected from N and O. In one embodiment, two R₁₁, takentogether with the adjacent atoms to which they are attached, formheteroaryl selected from pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl,pyranyl, thiopyranyl, diazinyl, thiazinyl, dioxinyl, and triazinyl, eachof which is optionally substituted as described herein (e.g., as in(XIV27)). In one embodiment, two R₁₁, taken together with the adjacentatoms to which they are attached, form 6-membered heteroaryl comprising1-2 nitrogen atoms. In one embodiment, two R₁₁, taken together with theadjacent atoms to which they are attached, form 6-membered heteroarylcomprising 1 nitrogen atom. In one embodiment, two R₁₁, taken togetherwith the adjacent atoms to which they are attached, form 6-memberedheteroaryl comprising 2 nitrogen atoms. In one embodiment, two R₁₁,taken together with the adjacent atoms to which they are attached, formheteroaryl selected from pyridinyl, pyridazinyl, and pyrimidinyl, eachof which is optionally substituted as described herein (e.g., as in(XIV27)).

(XIV33) In one embodiment, two R₁₁, taken together with the adjacentatoms to which they am attached, form 5- or 6-membered heterocyclylcomprising 1-3 heteroatoms selected from N, O, and S, optionallysubstituted as described herein (e.g., as in (XIV27)).

(XIV34) In one embodiment, two R₁₁, taken together with the adjacentatoms to which they are attached, form 5-membered heterocyclylcomprising 1-3 heteroatoms selected from N, O, and S, optionallysubstituted as described herein (e.g., as in (XIV27)). In oneembodiment, two R₁₁, taken together with the adjacent atoms to whichthey are attached, form 5-membered heterocyclyl comprising 1-2heteroatoms selected from N, O, and S. In one embodiment, two R₁₁, takentogether with the adjacent atoms to which they are attached, form5-membered heterocyclyl comprising 1-2 heteroatoms selected from N andO. In one embodiment, two R₁₁, taken together with the adjacent atoms towhich they are attached, form 5-membered heterocyclyl comprising 1heteroatom selected from N, O, and S. In one embodiment, two R₁₁, takentogether with the adjacent atoms to which they are attached, formheterocyclyl selected from pyrrolidinyl, tetrahydrofuranyl,tetrahydrothiophenyl, pyrazolidinyl, imidazolidinyl, oxazolidinyl,isoxazolidinyl, thiazolidinyl, isothiazolidinyl, triazolidinyl,oxadiazolidinyl, isoxadiazolidinyl, thiadiazolidinyl,isothiadiazolidinyl, dioxolanyl, and dithiolanyl, each of which isoptionally substituted as described herein (e.g., as in (XIV27)). In oneembodiment, the heterocycle is selected from pyrrolidin-2-onyl,pyrrolidin-3-onyl, pyrrolidine-2,3-dionyl pyrrolidine-2,5-dionyl, andimidazolidinonyl, oxazolidinonyl, and thiazolidinonyl

(XIV35) In one embodiment, two R₁₁, taken together with the adjacentatoms to which they are attached, form 6-membered heterocyclylcomprising 1-3 heteroatoms selected from N, O, and S, optionallysubstituted as described herein (e.g., as in (XIV27)). In oneembodiment, two R₁₁, taken together with the adjacent atoms to whichthey are attached, form 6-membered heterocyclyl comprising 1-2heteroatoms selected from N, O, and S. In one embodiment, two R₁₁, takentogether with the adjacent atoms to which they are attached, form6-membered heterocyclyl comprising 1-2 heteroatoms selected from N andO. In one embodiment, two R₁₁, taken together with the adjacent atoms towhich they are attached, form 6-membered heterocyclyl comprising 1heteroatom selected from N, O, and S. In one embodiment, two R₁₁, takentogether with the adjacent atoms to which they are attached, formheterocyclyl selected from dihydropyridinyl, tetrahydropyridinyl,piperidinyl, dihydropyrazinyl, tetrahydropyrazinyl, piperazinyl,morpholinyl, oxazinyl, dihydrooxazinyl, oxazinanyl, dihydrothiazinyl,tetrahydrothiazinyl, thiomorpholinyl, pyranyl, dihydropyranyl,tetrahydropyranyl, dioxinyl, dihydrodioxinyl, dioxanyl, thiopyranyl,dihydrothiopyranyl, tetrahydropyranyl, and dithianyl. In one embodiment,pyridinonyl, dihydropyridinonyl, hexahydropyridazinyl,hexahydropyrimidinyl, triazinanyl, piperidin-2-onyl, piperidin-3-onyl,piperidin-4-onyl, piperidine-2,6-dionyl, tetrahydropyrimidinonyl,oxazinanonyl, and thiazinanonyl, each of which is optionally substitutedas described herein (e.g., as in (XIV27)).

Any of the substituents described herein for any of A₁, A₂, X₁, X₂, X₃,X₄, X₅, X₆, X₇, X₈, L₁, R₀, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, R₉, R₁₀,R₁₁, m, n, and p can be combined with any of the substituents describedherein for one or more of the remainder of A₁, A₂, X₁, X₂, X₃, X₄, X₅,X₆, X₇, X₈, L₁, R₀, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, R₉, R₁₀, R₁₁, m, n,and p.

In one embodiment, a compound of Formula I is of Formula IIa, IIa′, IIb,IIb′, IIc, IIc′, IId, IId′, IIe, IIe′, IIf, IIg, IIg′, IIh, IIh′, IIi,IIi′, IIj, IIj′, IIk, or IIk′:

or a pharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof, wherein:

A₁, A₂, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, L₁, R₀, R₁, R₂, R₃, R₄, R₅, R₆,R₇, R₈, R₉, R₁₀, R₁₁, m, n, and p are each as defined in Formula I;

bis 0, 1, or 2; and

cis 1, 2, or 3.

In one embodiment, b is 0 or 1.

In one embodiment, b is 0.

In one embodiment, b is 1.

In one embodiment, b is 2.

In one embodiment, c is 1, 2, or 3, wherein 0, 1, 2, or 3 R₉ is H.

In one embodiment, c is 1, 2, or 3, wherein 0, 1, or 2 R₉ is H.

In one embodiment, c is 1, 2, or 3, wherein 0 or 1 R₉ is H.

In one embodiment, c is 1, 2, or 3, wherein 1, 2, or 3 R₉ is H.

In one embodiment, c is 1, 2, or 3, wherein 1 or 2 R₉ is H.

In one embodiment, c is 1, 2, or 3, wherein 2 or 3 R₉ is H.

In one embodiment, c is 1, 2, or 3, wherein 0 R₉ is H.

In one embodiment, c is 1, 2, or 3, wherein 1 R₉ is H.

In one embodiment, c is 1, 2, or 3, wherein 2 R₉ are H.

In one embodiment, c is 1, 2, or 3, wherein 3 R₉ are H.

Any of the substituents described herein for any of A₁, A₂, X₁, X₂, X₃,X₄, X₅, X₆, X₇, X₈, L₁, R₀, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, R₉, R₁₀,R₁₁, m, n, p, b, and c for example, in Formula I and any of Formula IIa,IIa′, IIb, IIb′, IIc, IIc′, IId, IId′, IIe, IIe′, IIf, IIg, IIg′, IIh,IIh′, IIi, IIi′, IIj, IIj′, IIk, or IIk′, as applicable, can be combinedwith any of the substituents described herein for one or more of theremainder of A₁, A₂, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, L₁, R₀, R₁, R₂, R₃,R₄, R₅, R₆, R₇, R₈, R₉, R₁₀, R₁₁, m, n, p, b, and c, for example, inFormula I and any of Formula IIa, IIa′, IIb, IIb′, IIc, IIc′, IId, IId′,IIe, IIe′, IIf, IIg, IIg′, IIh, IIh′, IIi, IIi′, IIj, IIj′, IIk, orIIk′, as applicable.

In one embodiment, a compound of Formula I is of Formula IIIa, IIIa′,IIIb, IIIb′, IIIc, IIIc′, IIId, IIId′, IIIe, IIIe′, IIIf, IIIf′, IIIg,IIIg′, IIIh, IIIh′, IIIi, IIIi′, IIIj, IIIj′, IIIk, or IIIk′:

or a pharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof, wherein:

-   -   A₂, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, L₁, R₀, R₁, R₂, R₃, R₄, R₅,        R₆, R₇, R₈, R₉, R₁₀, R₁₁, m, n, and p, are each as defined in        Formula I; and a is 0, 1, 2, 3, 4, or 5.

In one embodiment, a is 0, 1, 2, 3, or 4.

In one embodiment, a is 0, 1, 2, or 3.

In one embodiment, a is 0, 1, or 2.

In one embodiment, a is 0 or 1.

In one embodiment, a is 0.

In one embodiment, a is 1.

In one embodiment, a is 2.

In one embodiment, a is 3.

In one embodiment, a is 4.

In one embodiment, a is 5.

In one embodiment, L₁ is —C(O)—, —C(O)(CH₂)_(n)—, or —C(O)NH—.

In one embodiment, L₁ is —C(O)—.

In one embodiment, L₁ is —C(O)(CH₂)_(n)—.

In one embodiment, L₁ is —C(O)(CH₂)—.

In one embodiment, L₁ is —C(O)NH—.

Any of the substituents described herein for any of A₂, X₁, X₂, X₃, X₄,X₅, X₆, X₇, X₈, L₁, R₀, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, R₉, R₁₀, R₁₁, m,n, p, and a, for example, in Formula I and any of Formula IIIa, IIIa′,IIIb, IIIb′, IIIc, IIIc′, IIId, IIId′, IIIe, IIIe′, IIIf, IIIf′, IIIg,IIIg′, IIIh, IIIh′, IIIi, IIIi′, IIIj, IIIj′, IIIk, or IIIk′, asapplicable, can be combined with any of the substituents describedherein for one or more of the remainder of A₂, X₁, X₂, X₃, X₄, X₅, X₆,X₇, X₈, L₁, R₀, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, R₉, R₁₀, R₁₁, m, n, p,and a, for example, in Formula I and any of Formula IIIa, IIIa′, IIIb,IIIb′, IIIc, IIIc′, IIId, IIId′, IIIe, IIIe′, IIIf, IIIf′, IIIg, IIIg′,IIIh, IIIh′, IIIi, IIIi′, IIIj, IIIj′, IIIk, or IIIk′, as applicable.

In one embodiment, a compound of Formula I is of Formula IVa, IVa′, IVb,IVb′, IVc, IVc′, IVd, IVd′, IVe, IVe′, IVf, IVf′, IVg, IVg′, IVh, IVh′,IVi, IVi′, IVj, or IVj′:

or a pharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof, wherein:

A₂, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, R₀, R₁, R₄, R₅, R₆, R₇, R₈, R₉, R₁₀,R₁₁, and m are each as defined in Formula I; and a is 0, 1, 2, 3, 4, or5.

In one embodiment, a is 0, 1, 2, 3, or 4.

In one embodiment, a is 0, 1, 2, or 3.

In one embodiment, a is 0, 1, or 2.

In one embodiment, a is 0 or 1.

In one embodiment, a is 0.

In one embodiment, a is 1.

In one embodiment, a is 2.

In one embodiment, a is 3.

In one embodiment, a is 4.

In one embodiment, a is 5.

Any of the substituents described herein for any of A₂, X₁, X₂, X₃, X₄,X₅, X₆, X₇, X₈. R₀, R₁, R₄, R₅, R₆, R₇, R₈, R₉, R₁₀, R₁₁, m, and a, forexample, in Formula I and any of Formula IVa, IVa′, IVb, IVb′, IVc,IVc′, IVd, IVd′, IVe, IVe′, IVf, IVf′, IVg, IVg′, IVh, IVh′, IVi, IVi′,IVj, or IVj′, as applicable, can be combined with any of thesubstituents described herein for one or more of the remainder of A₂,X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, R₀, R₁, R₄, R₅, R₆, R₇, R₈, R₉, R₁₀,R₁₁, m, and a, for example, in Formula I and any of Formula IVa, IVa′,IVb, IVb′, IVc, IVc′, IVd, IVd′, IVe, IVe′, IVf, IVf′, IVg, IVg′, IVh,IVh′, IVi, IVi′, IVj, or IVj′, as applicable.

In one embodiment, a compound of Formula I is of Formula I-i:

or a pharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof, wherein:

A₂, X₈, R₀, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, R₉, R₁₀, R₁₁, and m are eachas defined in Formula I;

a is 0, 1, 2, 3, 4, or 5;

L₂ is a bond, —NH—, —N(C₁-C₆alkyl)-, or —CH₂—;

is a single or double bond;

Y is C when

is a double bond, CH when

is a single bond, or N;

Z is CH₂, CH(R₁₁), NH, N(R₁₁), O, or S; and

q is 1 or 2.

(a1) In one embodiment, a is 0.

(b1) In one embodiment, R₁ is H.

(c1) In one embodiment, R₂ is H.

(d1) In one embodiment, L₂ is —CH₂—.

(e1) In one embodiment, L₂ is —NH—.

(f1) In one embodiment, L₂ is a bond.

(g1) In one embodiment, R₃ is H.

(h1) In one embodiment, R₃ is OCH₃.

(i1) In one embodiment, R₄ is F.

(j1) In one embodiment, R₄ is F and is at the mesa position indicated inthe above Formula I-i.

(k1) In one embodiment, X₈ is N.

(l1) In one embodiment, X₈ is CR₉.

(m1) In one embodiment, X₈ is CR₉ and R₉ is H.

(n1) In one embodiment, Y is C.

(o1) In one embodiment, Y is CH.

(p1) In one embodiment, Y is N.

(q1) In one embodiment, Z is O.

(r1) In one embodiment, Y is N, Z is O, and g is 2.

(s1) In one embodiment, Z is N(R₁₁).

(t1) In one embodiment, R₁₁ is methyl.

(u1) In one embodiment, Y is N, Z is N(R₁₁), R₁₁ is methyl, and q is 2.

(v1) In one embodiment, Z is NH.

(w1) In one embodiment, Y is C,

is a double bond, Z is NH, and q is 2.

(x1) In one embodiment,

(y1) In one embodiment,

(z1) In one embodiment,

is at the para position relative to the position where L₂ is bonded.

(aa1) In one embodiment,

is at a meta position relative to the position where L₂ is bonded.

In one embodiment, a compound of Formula I is of Formula I-ii:

or a pharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof, wherein:

A₂, R₀, R₁, R₄, R₅, R₆, R₇, R₈, R₁₁, and m are each as defined inFormula I;

a is 0, 1, 2, 3, 4, or 5;

is a single or double bond;

Y is C when

is a double bond, CH when

is a single bond, or N;

Z is CH₂, CH(R₁₁), NH, N(R₁₁), O, or S; and

g is 1 or 2.

(a2) In one embodiment, a is 0.

(b2) In one embodiment, R₁ is H.

(c2) In one embodiment, R₄ is F.

(d2) In one embodiment, R₄ is F and is at the meta position indicated inthe above Formula I-ii.

(e2) In one embodiment. Y is C.

(f2) In one embodiment, Y is CH.

(g2) In one embodiment, Y is N.

(h2) In one embodiment, Z is O.

(i2) In one embodiment, Y is N, Z is O, and q is 2.

(j2) In one embodiment, Z is N(R₁₁).

(k2) In one embodiment, R₁₁ is methyl.

(l2) In one embodiment, Y is N, Z is N(R₁₁), a R₁₁ is methyl, and q is2.

(m2) In one embodiment, Z is NH.

(n2) In one embodiment, Y is C,

is a double bond, Z is —NH—, and q is 2.

(o2) In one embodiment, q is 1.

(p2) In one embodiment, q is 2.

(q2) In one embodiment,

is at the para position indicated in the above Formula I-ii.

(r2) In one embodiment,

is at a meta position indicated in the above Formula I-ii.

In one embodiment, a compound of Formula I is of Formula I-iii:

or a pharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof, wherein:

A₂, X₈, R₀, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, R₉, R₁₀, R₁₁, and m are eachas defined in Formula I;

a is 0, 1, 2, 3, 4, or 5;

L₂ is a bond, —NH—, —N(C₁-C₆alkyl)-, or —CH₂—;

f is C or N; and

each of b, c, d, and a is independently CH, C(R₁₁), N, NH, N(R₁₁), O, orS.

(a3) In one embodiment, a is 0.

(b3) In one embodiment, R₁ is H.

(c3) In one embodiment. R₂ is H.

(d3) In one embodiment, L₂ is —CH₂—.

(e3) In one embodiment, L₂ is —NH—.

(f3) In one embodiment, L₂ is a bond.

(g3) In one embodiment, R₃ is H.

(h3) In one embodiment, R₃ is OCH₃.

(i3) In one embodiment, R₄ is F.

(j3) In one embodiment, R₄ is F and is at the meta position indicated inthe above Formula i-iii.

(k3) In one embodiment, X₈ is N.

(l3) In one embodiment, X₈ is CR₉.

(m3) In one embodiment, X₈ is CR₉ and R₉ is H.

(n3) In one embodiment, f is N.

(o3) In one embodiment, b is CH.

(p3) In one embodiment, c is N.

(q3) In one embodiment, d is N.

(r3) In one embodiment, e is N.

(s3) In one embodiment, f is N, b is CH, and each of c, d, and e is N.

(t3) In one embodiment, f is C.

(u3) In one embodiment, b is CH.

(v3) In one embodiment, c is N(R₁₁).

(w3) In one embodiment, R₁₁ is methyl.

(x3) In one embodiment, c is NH.

(y3) In one embodiment, d is N.

(z3) In one embodiment, c is CH.

(aa3) In one embodiment, f is C, b is CH, c is N(R₁₁), R₁₁ is methyl, cis NH, d is N. and a is CH.

(bb3) In one embodiment,

is at the para position relative to the position where L₂ is bonded.

(cc3) In one embodiment,

is at a meta position relative to the position where L₂ is bonded.

In one embodiment, the compound is represented by Formula I-iv:

or a pharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof, wherein:

A₂, R₀, R₁, R₄, R₅, R₆, R₇, R₈, R₁₁, and m are each as defined inFormula I;

a is 0, 1, 2, 3, 4, or 5;

f is C or N; and

each of b, c, d, and e is independently CH, C(R₁₁), N, NH, N(R₁₁), O, orS.

(a4) In one embodiment, a is 0.

(b4) In one embodiment, R₁ is H.

(c4) In one embodiment, R₄ is F.

(d4) In one embodiment, R₄ is F and is at the meta position indicated inthe above Formula I-iv.

(e4) In one embodiment, f is N.

(f4) In one embodiment, b is CH.

(g4) In one embodiment, c is N.

(h4) In one embodiment, d is N.

(i4) In one embodiment, e is N.

(j4) In one embodiment, f is N, b is CH, and each of b, c, d, and e isN.

(k4) In one embodiment, f is C.

(l4) In one embodiment, b is CH.

(m4) In one embodiment, c is N(R₁₁).

(n4) In one embodiment, R₁₁ is methyl.

(o4) In one embodiment, c is NH.

(p4) In one embodiment, d is N.

(q4) In one embodiment, e is CH.

(r4) In one embodiment, f is C, b is CH, c is N(R₁₁), R₁₁ is methyl, cis NH, d is N, and a is CH.

(s4) In one embodiment,

is at the para position indicated in the above Formula I-iv.

(t4) In one embodiment,

is at a meta position indicated in the above Formula I-iv.

In one embodiment, the compound is represented by Formula I-v:

or a pharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof, wherein:

A₂, X₈, R₀, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, R₉, R₁₀, R₁₁, and m are eachas defined in Formula I;

a is 0, 1, 2, 3, 4, or 5;

L₂ is a bond, —NH—, —N(C₁-C₆alkyl)-, or —CH₂—; and

each of f, g, h, i, and j is independently CH, C(R₁₁), or N,

wherein:

-   -   at least one of f, g, h, and i is C(R₁₁); and    -   no more than two of f, g, h, i, and j is N.

(a5) In one embodiment, a is 0.

(b5) In one embodiment, R₁ is H.

(c5) In one embodiment, R₂ is H.

(d5) In one embodiment, L₂ is —CH₂—.

(e5) In one embodiment, L₂ is —NH—.

(f5) In one embodiment, L₂ is a bond.

(g5) In one embodiment, R₃ is H.

(h5) In one embodiment, R₃ is OCH₃.

(i5) In one embodiment, R₄ is F.

(j5) In one embodiment, R₄ is F is at the meta position indicated in theabove Formula I-v.

(k5) In one embodiment, X₈ is N.

(l5) In one embodiment, X₈ is CR₉.

(m5) In one embodiment, X₈ is CR₉ and R₉ is H.

(n5) In one embodiment, j is CH.

(o5) In one embodiment, i is CH.

(p5) In one embodiment, i is C(R₁₁).

(q5) In one embodiment, h is CH.

(r5) In one embodiment, h is C(R₁₁).

(s5) In one embodiment, g is CH.

(t5) In one embodiment, g is C(R₁₁).

(u5) In one embodiment, f is CH.

(v5) In one embodiment,

is at the para position relative to the position where L₂ is bonded.

(w5) In one embodiment,

is at a meta position relative to the position where L₂ is bonded.

In one embodiment, the compound is represented by Formula I-vi:

or a pharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof, wherein:

A₂, R₀, R₁, R₄, R₅, R₆, R₇, R₈, R₁₁, and m are each as defined inFormula I;

a is 0, 1, 2, 3, 4, or 5; and

each of f, g, h, i, and j is independently CH, C(R₁₁), or N,

wherein:

-   -   at least one of f, g, h, and i is C(R₁₁); and    -   no more than two of f, g, h, i, and j is N.

(a6) In one embodiment, a is 0.

(b6) In one embodiment, R₁ is H.

(c6) In one embodiment, R₂ is H.

(d6) In one embodiment, R₄ is F.

(e6) In one embodiment, R₄ is F and is at the meta position indicated inthe above Formula I-vi.

(f6) In one embodiment, j is CH.

(g6) In one embodiment, i is CH.

(h6) In one embodiment, i is C(R₁₁).

(i6) In one embodiment, h is CH.

(j6) In one embodiment, h is C(R₁₁)

(k6) In one embodiment, g is CH.

(l6) In one embodiment, g is C(R₁₁).

(m6) In one embodiment, f is CH.

(n6) In one embodiment,

is at the para position indicated in the above Formula I-vi.

(o6) In one embodiment,

is at a meta position indicated in the above Formula I-vi.

Non-limiting illustrative compounds of the application are included inTable A:

TABLE A Compound ID Structure I-1

I-2

I-3

I-4

I-5

I-6

I-7

I-8

I-9

I-10

I-11

I-12

I-13

I-14

I-15

I-16

I-17

I-18

A compound that binds to an allosteric site in EGFR, such as thecompounds of the present application (e.g., the compounds of theformulae disclosed herein), optionally in combination with a secondagent that prevents EGFR dimer formation, are capable of modulating(e.g., inhibiting or decreasing) EGFR activity. In some embodiments, thecompounds of the present application are capable of inhibiting ordecreasing EGFR activity, without a second agent (e.g., an antibody suchas cetuximab, trastuzumab, or panitumumab). In other embodiments, thecompounds of the present application, in combination with a second agentthat prevents EGFR dimer formation (e.g., an antibody such as cetuximab,trastuzumab, or panitumumab), am capable of inhibiting or decreasingEGFR activity. In some embodiments, the second agent that prevents EGFRdimer formation is an antibody. In further embodiments, the second agentthat prevents EGFR dimer formation is cetuximab, trastuzumab, orpanitumumab. In further embodiments, the second agent that prevents EGFRdimer formation is cetuximab.

In some embodiments, the compounds of the present application arecapable of modulating (e.g., inhibiting or decreasing) the activity ofEGFR containing one or more mutations. In some embodiments, the mutantEGFR contains one or more mutations selected from T790M, L718Q, L844V,V948R. L858R, I941R. C797S, Del (e.g., deletion in exon 19), andInsertion (e.g., insertion in exon 20). In some embodiments, the mutantEGFR contains C797S. In other embodiments, the mutant EGFR contains acombination of mutations, wherein the combination is selected fromDel/L718Q, Del/L844V, Del/T790M, Del/T790M/L718Q, Del/T790M/L844V,L858R/L718Q, L858R/L844V, L858R/T790M, L858R/T790M/I941R, Del/T790M,Del/T790M/C797S, L858R/T790M/C797S, and L858R/T790M/L718Q. In otherembodiments, the mutant EGFR contains a combination of mutations,wherein the combination is selected from Del/L844V, L858R/L844V,L858R/T790M, L858R/T790M/I941R, L858R/T790M/C797S, Del/T790M. andDel/T790M/C797S. In other embodiments, the mutant EGFR contains acombination of mutations, wherein the combination is selected fromL858R/T790M, L858R/T790M/1941R, L858R/T790M/C797S, Del/T790M,Del/T790M/C797S, and L858R/T790M.

In some embodiments, the compounds of the present application incombination with a second agent that prevents EGFR dimer formation arecapable of modulating (e.g., inhibiting or decreasing) the activity ofEGFR containing one or more mutations (e.g., the EGFR containing one ormore mutations described herein). In some embodiments, the second agentthat prevents EGFR dimer formation is an antibody. In furtherembodiments, the second agent that prevents EGFR dimer formation iscetuximab, trastuzumab, or panitumumab. In further embodiments, thesecond agent that prevents EGFR dimer formation is cetuximab.

In some embodiments, the compounds of the present application arecapable of modulating (e.g., inhibiting or decreasing) the activity ofEGFR containing one or more mutations, but do not affect the activity ofa wild-type EGFR.

In other embodiments, the compounds of the present application incombination with a second agent that prevents EGFR dimer formation arecapable of modulating (e.g., inhibiting or decreasing) the activity ofEGFR containing one or more mutations, but do not affect the activity ofa wild-type EGFR. In some embodiments, the second agent that preventsEGFR dimer formation is an antibody. In further embodiments, the secondagent that prevents EGFR dimer formation is cetuximab, trastuzumab, orpanitumumab. In further embodiments, the second agent that prevents EGFRdimer formation is cetuximab.

Modulation of EGFR containing one or more mutations, such as thosedescribed herein, but not a wild-type EGFR provides a novel approach tothe treatment, prevention, or amelioration of diseases including, butnot limited to, cancer and metastasis, inflammation, arthritis, systemiclupus erthematosus, skin-related disorders, pulmonary disorders,cardiovascular disease, ischemia, neurodegenerative disorders, liverdisease, gastrointestinal disorders, viral and bacterial infections,central nervous system disorders, Alzheimer's disease, Parkinson'sdisease, Huntington's disease, amyotrophic lateral sclerosis, spinalcord injury, and peripheral neuropathy.

In some embodiments, the compounds of the application exhibit greaterinhibition of EGFR containing one or more mutations as described hereinrelative to a wild-type EGFR. In certain embodiments, the compounds ofthe application exhibit at least 2-fold, 3-fold, 5-fold, 10-fold,25-fold, 50-fold or 100-fold greater inhibition of EGFR containing oneor more mutations as described herein relative to a wild-type EGFR. Invarious embodiments, the compounds of the application exhibit up to1000-fold greater inhibition of EGFR containing one or more mutations asdescribed herein relative to a wild-type EGFR. In various embodiments,the compounds of the application exhibit up to 10000-fold greaterinhibition of EGFR having a combination of mutations described hereinrelative to a wild-type EGFR.

In other embodiments, the compounds of the application in combinationwith a second agent that prevents EGFR dimer formation exhibit greaterinhibition of EGFR containing one or more mutations as described hereinrelative to a wild-type EGFR. In certain embodiments, the compounds ofthe application in combination with a second agent that prevents EGFRdimer formation exhibit at least 2-fold, 3-fold, 5-fold, 10-fold,25-fold, 50-fold or 100-fold greater inhibition of EGFR containing oneor more mutations as described herein relative to a wild-type EGFR. Invarious embodiments, the compounds of the application in combinationwith a second agent that prevents EGFR dimer formation exhibit up to1000-fold greater inhibition of EGFR containing one or more mutations asdescribed herein relative to a wild-type EGFR. In various embodiments,the compounds of the application in combination with a second agent thatprevents EGFR dimer formation exhibit up to 10000-fold greaterinhibition of EGFR having a combination of mutations described hereinrelative to a wild-type EGFR. In some embodiments, the second agent thatprevents EGFR dimer formation is an antibody. In further embodiments,the second agent that prevents EGFR dimer formation is cetuximab,trastuzumab, or panitumumab. In further embodiments, the second agentthat prevents EGFR dimer formation is cetuximab.

In some embodiments, the compounds of the application exhibit from about2-fold to about 10-fold greater inhibition of EGFR containing one ormore mutations as described herein relative to a wild-type EGFR. Invarious embodiments, the compounds of the application exhibit from about10-fold to about 100-fold greater inhibition of EGFR containing one ormore mutations as described herein relative to a wild-type EGFR. Invarious embodiments, the compounds of the application exhibit from about100-fold to about 1000-fold greater inhibition of EGFR containing one ormore mutations as described herein relative to a wild-type EGFR. Invarious embodiments, the compounds of the application exhibit from about1000-fold to about 10000-fold greater inhibition of EGFR containing oneor more mutations as described herein relative to a wild-type EGFR.

In other embodiments, the compounds of the application in combinationwith a second agent that prevents EGFR dimer formation exhibit fromabout 2-fold to about 10-fold greater inhibition of EGFR containing oneor more mutations as described herein relative to a wild-type EGFR. Inother embodiments, the compounds of the application in combination witha second agent that prevents EGFR dimer formation exhibit from about10-fold to about 100-fold greater inhibition of EGFR containing one ormore mutations as described herein relative to a wild-type EGFR. Inother embodiments, the compounds of the application in combination witha second agent that prevents EGFR dimer formation exhibit from about100-fold to about 1000-fold greater inhibition of EGFR containing one ormore mutations as described herein relative to a wild-type EGFR. Inother embodiments, the compounds of the application in combination witha second agent that prevents EGFR dimer formation exhibit from about1000-fold to about 10000-fold greater inhibition of EGFR containing oneor more mutations as described herein relative to a wild-type EGFR. Inother embodiments, the second agent that prevents EGFR dimer formationis an antibody. In further embodiments, the second agent that preventsEGFR dimer formation is cetuximab, trastuzumab, or panitumumab. Infurther embodiments, the second agent that prevents EGFR dimer formationis cetuximab.

In some embodiments, the inhibition of EGFR activity is measured byIC₅₀.

In some embodiments, the inhibition of EGFR activity is measured byEC₅₀.

In some embodiments, the compounds of the application bind to anallosteric site in EGFR. In some embodiments, the compounds of theapplication interact with at least one amino acid residue of EGFRselected from Lys745, Leu788, and Ala 743. In other embodiments, thecompounds of the application interact with at least one amino acidresidue of EGFR selected from Cys755, Leu777, Phe856, and Asp855. Inother embodiments, the compounds of the application interact with atleast one amino acid residue of EGFR selected from Met766, Ile759,Glu762, and Ala763. In other embodiments, the compounds of theapplication interact with at least one amino acid residue of EGFRselected from Lys745, Leu788, and Ala 743, at least one amino acidresidue of EGFR selected from Cys755, Leu777, Phe856, and Asp855, and atleast one amino acid residue of EGFR selected from Met766, Ile759,Glu762, and Ala763. In other embodiments, the compounds of theapplication do not interact with the any of the amino acid residues ofEGFR selected from Met793, Gly796, and Cys797.

In some embodiments, the application provides a compound (e.g., anallosteric kinase inhibitor), wherein the compound is a more potentinhibitor of a drug-resistant EGFR mutant relative to a wild type EGFR.For example, the compound can be at least about 2-fold, 3-fold, 5-fold,10-fold, 25-fold, 50-fold or about 100-fold more potent at inhibitingthe kinase activity of the drug-resistant EGFR mutant relative to awild-type EGFR. In some embodiments, the drug-resistant EGFR mutant isresistant to one or more known EGFR inhibitors, including but notlimited to gefitinib, erlotinib, afatinib, lapatinib, neratinib.

In some embodiments, the drug-resistant EGFR mutant comprises asensitizing mutation, such as Del and L858R.

In some embodiments, the application provides a compound (e.g., anallosteric kinase inhibitor) in combination with a second agent thatprevents EGFR dimer formation, wherein the compound is a more potentinhibitor of a drug-resistant EGFR mutant relative to a wild type EGFR.For example, the compound in combination with a second agent thatprevents EGFR dimer formation can be at least about 2-fold, 3-fold,5-fold, 10-fold, 25-fold, 50-fold or about 100-fold more potent atinhibiting the kinase activity of the drug-resistant EGFR mutantrelative to a wild-type EGFR. In some embodiments, the drug-resistantEGFR mutant is resistant to one or more known EGFR inhibitors, includingbut not limited to gefitinib, erlotinib, afatinib, lapatinib, neratinib,WZ4002, CL-387785, AZD9291, and CO-1686. In some embodiments, thedrug-resistant EGFR mutant comprises a sensitizing mutation, such as Deland L858R. In some embodiments, the second agent that prevents EGFRdimer formation is an antibody. In further embodiments, the second agentthat prevents EGFR dimer formation is cetuximab, trastuzumab, orpanitumumab. In further embodiments, the second agent that prevents EGFRdimer formation is cetuximab.

In some embodiments, the application provides a compound (e.g., anallosteric kinase inhibitor), wherein the compound inhibits kinaseactivity of a drug-resistant EGFR mutant harboring a sensitizingmutation (e.g., Del and L858R) and a drug-resistance mutation (e.g.,T790M, L718Q, C797S, and L844V) with less than a 10-fold difference inpotency (e.g., as measured by IC₅₀) relative to an EGFR mutant harboringthe sensitizing mutation but not the drug-resistance mutation. In someembodiments, the difference in potency is less than about 9-fold,8-fold, 7-fold, 6-fold, 5-fold, 4-fold, 3-fold, or 2-fold.

In other embodiments, the application provides a compound (e.g., anallosteric kinase inhibitor) in combination with a second agent thatprevents EGFR dimer formation, wherein the compound in combination withthe second agent inhibits kinase activity of a drug-resistant EGFRmutant harboring a sensitizing mutation (e.g., Del and L858R) and adrug-resistance mutation (e.g., T790M, L718Q, C797S, and L844V) withless than a 10-fold difference in potency (e.g., as measured by IC₅₀)relative to an EGFR mutant harboring the sensitizing mutation but notthe drug-resistance mutation. In some embodiments, the difference inpotency is less than about 9-fold, 8-fold, 7-fold, 6-fold, 5-fold,4-fold, 3-fold, or 2-fold. In some embodiments, the second agent thatprevents EGFR dimer formation is an antibody. In further embodiments,the second agent that prevents EGFR dimer formation is cetuximab,trastuzumab, or panitumumab. In further embodiments, the second agentthat prevents EGFR dimer formation is cetuximab.

In some embodiments, the application provides a compound (e.g., anallosteric kinase inhibitor), wherein the compound is more potent thanone or more known EGFR inhibitors, including but not limited togefitinib, erlotinib, afatinib, lapatinib, neratinib, WZ4002, CL-387785,AZD9291, and CO-1686, at inhibiting the activity of EGFR containing oneor more mutations as described herein. For example, the compound can beat least about 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold orabout 100-fold more potent (e.g., as measured by IC₅₀) than gefitinib,erlotinib, afatinib, lapatinib, neratinib, WZ4002, CL-387785, AZD9291,and CO-1686 at inhibiting the activity of the EGFR containing one ormore mutations as described herein.

In other embodiments, the application provides a compound (e.g., anallosteric kinase inhibitor) in combination with a second agent thatprevents EGFR dimer formation, wherein the compound in combination withthe second agent is more potent than one or more known EGFR inhibitors,including but not limited to gefitinib, erlotinib, afatinib, lapatinib,neratinib, WZ4002, CL-387785, AZD9291, and CO-1686, at inhibiting theactivity of EGFR containing one or more mutations as described herein,such as T790M, L718Q, L844V, L858R, C797S, and Del. For example, thecompound in combination with a second agent that prevents EGFR dimerformation can be at least about 2-fold, 3-fold, 5-fold, 10-fold,25-fold, 50-fold or about 100-fold more potent (e.g., as measured byIC₅₀) than gefitinib, erlotinib, afatinib, lapatinib, neratinib, WZ4002,CL-387785, AZD9291, and CO-1686 at inhibiting the activity of the EGFRcontaining one or more mutations as described herein. In someembodiments, the second agent that prevents EGFR dimer formation is anantibody. In further embodiments, the second agent that prevents EGFRdimer formation is cetuximab, trastuzumab, or panitumumab. In furtherembodiments, the second agent that prevents EGFR dimer formation iscetuximab.

In some embodiments, the application provides a compound (e.g., anallosteric kinase inhibitor), wherein the compound is less potent thanone or more known EGFR inhibitors, including but not limited togefitinib, erlotinib, afatinib, lapatinib, neratinib, WZ4002, CL-387785,AZD9291, and CO-1686, at inhibiting the activity of a wild-type EGFR.For example, the compound can be at least about 2-fold, 3-fold, 5-fold,10-fold, 25-fold, 50-fold or about 100-fold less potent (e.g., asmeasured by IC₅₀) than gefitinib, erlotinib, afatinib, lapatinib,neratinib, WZ4002, CL-387785, AZD9291, and CO-1686, at inhibiting theactivity of a wild-type EGFR.

In other embodiments, the application provides a compound (e.g., anallosteric kinase inhibitor) in combination with a second agent thatprevents EGFR dimer formation, wherein the compound in combination withthe second agent is less potent than one or more known EGFR inhibitors,including but not limited to gefitinib, erlotinib, afatinib, lapatinib,neratinib, WZ4002, CL-387785, AZD9291, and CO-1686, at inhibiting theactivity of a wild-type EGFR. For example, the compound in combinationwith a second agent that prevents EGFR dimer formation can be at leastabout 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold or about100-fold less potent (e.g., as measured by IC₅₀) than gefitinib,erlotinib, afatinib, lapatinib, neratinib, WZ4002, CL-387785, AZD9291,and CO-1686, at inhibiting the activity of a wild-type EGFR. In someembodiments, the second agent that prevents EGFR dimer formation is anantibody. In further embodiments, the second agent that prevents EGFRdimer formation is cetuximab, trastuzumab, or panitumumab. In furtherembodiments, the second agent that prevents EGFR dimer formation iscetuximab.

Potency of the inhibitor can be determined by EC₅₀ value. A compoundwith a lower EC₅₀ value, as determined under substantially similarconditions, is a more potent inhibitor relative to a compound with ahigher EC₅₀ value. In some embodiments, the substantially similarconditions comprise determining an EGFR-dependent phosphorylation level,in vitro or in vivo (e.g., in 3D cells expressing a wild type EGFR, amutant EGFR, or a fragment of any thereof).

Potency of the inhibitor can also be determined by IC₅₀ value. Acompound with a lower IC₅₀ value, as determined under substantiallysimilar conditions, is a more potent inhibitor relative to a compoundwith a higher IC₅₀ value. In some embodiments, the substantially similarconditions comprise determining an EGFR-dependent phosphorylation level,in vitro or in vivo (e.g., in 3D cells expressing a wild type EGFR, amutant EGFR, or a fragment of any thereof).

An EGFR sensitizing mutation comprises without limitation L858R, G719S,G719C, G719A, L861 Q, a deletion in exon 19 and/or an insertion in exon20. A drug-resistant EGFR mutant can have without limitation a drugresistance mutation comprising T790M, T854A, L718Q, C797S, or D761Y.

The selectivity between wild-type EGFR and EGFR containing one or moremutations as described herein can also be measured using cellularproliferation assays where cell proliferation is dependent on kinaseactivity. For example, murine Ba/F3 cells transfected with a suitableversion of wild-type EGFR (such as VIII; containing a WT EGFR kinasedomain), or Ba/F3 cells transfected with L858R/T790M, Del/T790M/L718Q,L858R/T790M/L718Q, L858R/T790M/C797S, Del/T790M/C797S,L858R/T790M/I941R, or Exon 19 deletion/T790M can be used. Proliferationassays are performed at a range of inhibitor concentrations (e.g., 10μM, 3 μM, 1.1 μM, 330 nM, 110 nM, 33 nM, 11 nM, 3 nM, 1 nM) and an EC₅₀is calculated.

An alternative method to measure effects on EGFR activity is to assayEGFR phosphorylation. Wild type or mutant (L858R/T790M. Del/T790M,Del/T790M/L718Q, L858R/T790M/C797S, Del/T790M/C797S, L858R/T790M/I941R,or L858R/T790M/L718Q) EGFR can be transfected into cells which do notnormally express endogenous EGFR and the ability of the inhibitor (usingconcentrations as above) to inhibit EGFR phosphorylation can be assayed.Cells are exposed to increasing concentrations of inhibitor andstimulated with EGF. The effects on EGFR phosphorylation are assayed byWestern Blotting using phospho-specific EGFR antibodies.

In another aspect, the present application relates to a compound thatbinds to an allosteric site in EGFR, wherein the compound exhibitsgreater than 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold,100-fold, or 1000-fold inhibition of EGFR containing one or moremutations as described herein (e.g., L858R/T790M, Del/T790M,Del/T790M/L718Q, L858R/T790M/C797S, Del/T790M/C797S, L858R/1790M/I941R,or L858R/I790M/L718Q) relative to a wild-type EGFR.

In other embodiments, the application provides a compound that binds toan allosteric site in EGFR in combination with a second agent thatprevents EGFR dimer formation, wherein the compound in combination withthe second agent greater than 2-fold, 3-fold, 5-fold, 10-fold, 25-fold,50-fold, 100-fold, or 1000-fold inhibition of EGFR containing one ormore mutations as described herein (e.g., L858R/T790M, Del/T790M,Del/T790M/L718Q, Del/T790M/C797S, L858R/T790M/C797S, L858R/T790M/I941R,or L858R/T790M/L718Q) relative to a wild-type EGFR. In some embodiments,the second agent that prevents EGFR dimer formation is an antibody. Infurther embodiments, the second agent that prevents EGFR dimer formationis cetuximab, trastuzumab, or panitumumab. In further embodiments, thesecond agent that prevents EGFR dimer formation is cetuximab.

Another aspect is an isotopically labeled compound of any of theformulae delineated herein. Such compounds have one or more isotopeatoms which may or may not be radioactive (e.g., ³H, ²H, ¹⁴C, ¹³C, ¹⁸F,³⁵S, ³²P, ¹²⁵I, and ¹³¹I) introduced into the compound. Such compoundsare useful for drug metabolism studies and diagnostics, as well astherapeutic applications.

The application also provides for a pharmaceutical compositioncomprising a compound disclosed herein, or a pharmaceutically acceptablesalt, hydrate, solvate, or stereoisomer thereof, together with apharmaceutically acceptable carrier.

In another aspect, the application provides a kit comprising a compoundcapable of inhibiting EGFR activity selected from one or more compoundsof disclosed herein, or a pharmaceutically acceptable salt, hydrate,solvate, or stereoisomer thereof, optionally in combination with asecond agent that prevents EGFR dimer formation and instructions for usein treating cancer.

In another aspect, the application provides a method of synthesizing acompound disclosed herein. The synthesis of the compounds of theapplication can be found herein and in the schemes and Examples below.Other embodiments are a method of making a compound of any of theformulae herein using any one, or combination of, reactions delineatedherein. The method can include the use of one or more intermediates orchemical reagents delineated herein.

The compounds of the application are defined herein by their chemicalstructures and/or chemical names. Where a compound is referred to byboth a chemical structure and a chemical name, and the chemicalstructure and chemical name conflict, the chemical structure isdeterminative of the compound's identity.

The recitation of a listing of chemical groups in any definition of avariable herein includes definitions of that variable as any singlegroup or combination of listed groups. The recitation of an embodimentfor a variable herein includes that embodiment as any single embodimentor in combination with any other embodiments or portions thereof.

Definitions

Listed below are definitions of various terms used to describe thisapplication. These definitions apply to the terms as they are usedthroughout this specification and claims, unless otherwise limited inspecific instances, either individually or as part of a larger group.

The term “alkyl,” as used herein, refers to saturated, straight- orbranched-chain hydrocarbon radicals containing, in certain embodiments,between one and six, or one and eight carbon atoms, respectively.Examples of C₁-C₆ alkyl radicals include, but are not limited to,methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, neopentyl,n-hexyl radicals; and examples of C₁-C₈ alkyl radicals include, but arenot limited to, methyl, ethyl, propyl, isopropyl, n-butyl, tert-butylneopentyl, n-hexyl, heptyl, octyl radicals.

The term “alkenyl,” as used herein, denotes a monovalent group derivedfrom a hydrocarbon moiety containing, in certain embodiments, from twoto six, or two to eight carbon atoms having at least one carbon-carbondouble bond. The double bond may or may not be the point of attachmentto another group. Alkenyl groups include, but are not limited to, forexample, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, heptenyl,octenyl and the like.

The term “alkynyl,” as used herein, denotes a monovalent group derivedfrom a hydrocarbon moiety containing, in certain embodiments, from twoto six, or two to eight carbon atoms having at least one carbon-carbontriple bond. The alkynyl group may or may not be the point of attachmentto another group. Representative alkynyl groups include, but are notlimited to, for example, ethynyl, 1-propynyl, 1-butynyl, heptynyl,octynyl and the like.

The term “alkoxy” refers to an —O-alkyl radical.

The term “aryl,” as used herein, refers to a mono- or poly-cycliccarbocyclic ring system having one or more aromatic rings, fused ornon-fused, including, but not limited to, phenyl, naphthyl,tetrahydronaphthyl, indanyl, indenyl and the like.

The term “aralkyl,” as used herein, refers to an alkyl residue attachedto an aryl ring. Examples include, but are not limited to, benzyl,phenethyl and the like.

The term “cycloalkyl,” as used herein, denotes a monovalent groupderived from a monocyclic or polycyclic saturated or partiallyunsaturated carbocyclic ring compound. Examples of C₃-C₈ cycloalkylinclude, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cyclopentyl and cyclooctyl; and examples ofC₃-C₁₂-cycloalkyl include, but not limited to, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, bicyclo [2.2.1] heptyl, and bicyclo [2.2.2]octyl. Also contemplated is a monovalent group derived from a monocyclicor polycyclic carbocyclic ring compound having at least onecarbon-carbon double bond by the removal of a single hydrogen atom.Examples of such groups include, but are not limited to, cyclopropenyl,cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl,and the like.

The term “hetcroaryl,” as used herein, refers to a mono- or poly-cyclic(e.g., bi-, or tri cyclic or more) fused or non-fused, radical or ringsystem having at least one aromatic ring, having from five to ten ringatoms of which one ring atoms is selected from S, O, and N; zero, one,or two ring atoms are additional heteroatoms independently selected fromS, O, and N; and the remaining ring atoms are carbon. Heteroarylincludes, but is not limited to, pyridinyl, pyrazinyl, pyrimidinyl,pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isooxazolyl,thiadiazolyl, oxadiazolyl, thiophenyl, furanyl, quinolinyl,isoquinolinyl, benzimidazolyl, benzooxazolyl, quinoxalinyl, and thelike.

The term “heteroaralkyl,” as used herein, refers to an alkyl residueattached to a heteroaryl ring. Examples include, but are not limited to,pyridinylmethyl, pyrimidinylethyl and the like.

The term “heterocyclyl,” or “heterocycloalkyl,” as used herein, refersto a non-aromatic 3-, 4-, 5-, 6- or 7-membered ring or a bi- ortri-cyclic group fused of non-fused system, where (i) each ring containsbetween one and three heteroatoms independently selected from oxygen,sulfur and nitrogen, (ii) each 5-membered ring has 0 to 1 double bondsand each 6-membered ring has 0 to 2 double bonds, (iii) the nitrogen andsulfur heteroatoms may optionally be oxidized, and (iv) the nitrogenheteroatom may optionally be quaternized. Representativeheterocycloalkyl groups include, but are not limited to, [1,3]dioxolane,pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl,piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl,thiazolidinyl, isothiazolidinyl, and tetrahydrofuryl.

The term “alkylamino” refers to a group having the structure —NH(C₁-C₁₂alkyl), e.g., —NH(C₁-C₆ alkyl), where C₁-C₁₂ alkyl is as previouslydefined.

The term “dialkylamino” refers to a group having the structure —N(C₁-C₁₂alkyl)₂, e.g., —NH(C₁-C₆ alkyl), where C₁-C₁₂ alkyl is as previouslydefined.

The term “acyl” includes residues derived from acids, including but notlimited to carboxylic acids, carbamic acids, carbonic acids, sulfonicacids, and phosphorous acids. Examples include aliphatic carbonyls,aromatic carbonyls, aliphatic sulfonyls, aromatic sulfinyls, aliphaticsulfinyls, aromatic phosphates and aliphatic phosphates. Examples ofaliphatic carbonyls include, but are not limited to, acetyl, propionyl,2-fluoroacetyl, butyryl, 2-hydroxy acetyl, and the like.

In accordance with the application, any of the aryls, substituted aryls,heteroaryls and substituted heteroaryls described herein, can be anyaromatic group. Aromatic groups can be substituted or unsubstituted.

The terms “hal,” “halo,” and “halogen,” as used herein, refer to an atomselected from fluorine, chlorine, bromine and iodine.

As described herein, compounds of the application may optionally besubstituted with one or more substituents, such as are illustratedgenerally above, or as exemplified by particular classes, subclasses,and species of the application. It will be appreciated that the phrase“optionally substituted” is used interchangeably with the phrase“substituted or unsubstituted.” In general, the term “substituted”,whether preceded by the term “optionally” or not, refers to thereplacement of hydrogen radicals in a given structure with the radicalof a specified substituent. Unless otherwise indicated, an optionallysubstituted group may have a substituent at each substitutable positionof the group, and when more than one position in any given structure maybe substituted with more than one substituent selected from a specifiedgroup, the substituent may be either the same or different at everyposition. The terms “optionally substituted”, “optionally substitutedalkyl,” “optionally substituted “optionally substituted alkenyl,”“optionally substituted alkynyl”, “optionally substituted cycloalkyl,”“optionally substituted cycloalkenyl,” “optionally substituted aryl”,“optionally substituted heteroaryl,” “optionally substituted aralkyl”,“optionally substituted heteroaralkyl,” “optionally substitutedheterocycloalkyl,” and any other optionally substituted group as usedherein, refer to groups that are substituted or unsubstituted byindependent replacement of one, two, or three or more of the hydrogenatoms thereon with substituents including, but not limited to:

—F, —Cl, —Br, —I, —OH, protected hydroxy, —NO₂, —CN, —NH₂, protectedamino, —NH—C₁-C₁₂-alkyl, —NH—C₂-C₁₂-alkenyl, —NH—C₂-C₁₂-alkenyl,—NH—C₃-C₁₂-cycloalkyl, —NH-aryl, —NH-heteroaryl, —NH-heterocycloalkyl,-dialkylamino, -diarylamino, -diheteroarylamino, —O—C₁-C₁₂-alkyl,—O—C₂-C₁₂-alkenyl, —O—C₂-C₁₂-alkenyl, —O—C3-C₁₂-cycloalkyl, —O-aryl,—O-heteroaryl, —O-heterocycloalkyl, —C(O)—C₁-C₁₂-alkyl,—C(O)—C₂-C₁₂-alkenyl, —C(O)—C₂-C₁₂-alkenyl, —C(O)—C₃-C₁₂-cycloalkyl,—C(O)-aryl, —C(O)— heteroaryl,

˜C(O)-heterocycloalkyl, —CONH₂, —CONH—C₁-C₁₂-alkyl,—CONH—C₂-C₁₂-alkenyl, —CONH—C₂-C₁₂-alkenyl, —CONH—C₃-C₁₂-cycloalkyl,—CONH-aryl, —CONH-heteroaryl, —CONH-heterocycloalkyl,—OCO₂—C₁-C₁₂-alkyl, —OCO₂—C₂-C₁₂-alkenyl, —OCO₂—C₂-C₁₂-alkenyl,—OCO₂—C₃-C₁₂-cycloalkyl, —OCO₂-aryl, —OCO₂-heteroaryl,—OCO₂-heterocycloalkyl, —OCONH₂,—OCONH—C₁-C₁₂-alkyl, —OCONH—C₂-C₁₂-alkenyl, —OCONH—C₂-C₁₂-alkenyl,—OCONH—C₃-C₁₂-cycloalkyl, —OCONH-aryl, —OCONH-heteroaryl, —OCONH—heterocycloalkyl,—NHC(O)—C₁-C₁₂-alkyl, —NHC(O)—C₂-C₁₂-alkenyl, —NHC(O)—C₂-C₁₂-alkenyl,—NHC(O)—C₃-C₁₂-cycloalkyl, —NHC(O)-aryl, —NHC(O)-heteroaryl, —NHC(O)—heterocycloalkyl,—NHCO₂—C₁-C₁₂-alkyl, —NHCO₂—C₂-C₁₂-alkenyl, —NHCO₂—C₂-C₁₂-alkenyl,—NHCO₂—C₃-C₁₂-cycloalkyl, —NHCO₂-aryl, —NHCO₂-heteroaryl,—NHCO₂-heterocycloalkyl, NHC(O)NH₂, —NHC(O)NH—C₁-C₁₂-alkyl,—NHC(O)NH—C₂-C₁₂-alkenyl,—NHC(O)NH—C₂-C₁₂-alkenyl, —NHC(O)NH—C₃-C₁₂-cycloalkyl, —NHC(O)NH-aryl,—NHC(O)NH-heteroaryl, NHC(O)NH-heterocycloalkyl, —NHC(S)NH₂,—NHC(S)NH—C₁-C₁₂-alkyl, —NHC(S)NH—C₂-C₁₂-alkenyl,—NHC(S)NH—C₂-C₁₂-alkenyl, —NHC(S)NH—C₃-C₁₂-cycloalkyl, —NHC(S)NH-aryl,—NHC(S)NH-heteroaryl, —NHC(S)NH-heterocycloalkyl, —NHC(NH)NH₂,—NHC(NH)NH—C₁-C₁₂-alkyl, —NHC(NH)NH—C₂-C₁₂-alkenyl,—NHC(NH)NH—C₂-C₁₂-alkenyl, —NHC(NH)NH—C₃-C₁₂-cycloalkyl,—NHC(NH)NH-aryl, —NHC(NH)NH-heteroaryl, —NHC(NH)NHheterocycloalkyl,—NHC(NH)—C₁-C₁₂-alkyl, —NHC(NH)—C₂-C₁₂-alkenyl, —NHC(NH)—C₂-C₁₂-alkenyl,—NHC(NH)—C₃-C₁₂-cycloalkyl, —NHC(NH)-aryl, —NHC(NH)-heteroaryl,—NHC(NH)-heterocycloalkyl, —C(NH)NH—C₁-C₁₂-alkyl,—C(NH)NH—C₂-C₁₂-alkenyl, —C(NH)NH—C₂-C₁₂-alkenyl,C(NH)NH—C₃-C₁₂-cycloalkyl, —C(NH)NH-aryl, —C(NH)NH-heteroaryl,—C(NH)NHheterocycloalkyl, —S(O)—C₁-C₁₂-alkyl, —S(O)—C₂-C₁₂-alkenyl,—S(O)—C₂-C₁₂-alkenyl, —S(O)—C₃-C₁₂-cycloalkyl, —S(O)-aryl,—S(O)-heteroaryl, —S(O)-heterocycloalkyl —SO₂NH₂, —SO₂NH—C₁-C₁₂-alkyl,—SO₂NH—C₂-C₁₂-alkenyl, —SO₂NH—C₂-C₁₂-alkenyl, —SO₂NH—C₃-C₁₂-cycloalkyl,—SO₂NH-aryl, —SO₂NH-heteroaryl, —SO₂NH-heterocycloalkyl,—NHSO₂—C₁-C₁₂-alkyl, —NHSO₂—C₂-C₁₂-alkenyl, —NHSO₂—C₂-C₁₂-alkenyl,—NHSO₂—C₃-C₁₂-cycloalkyl, —NHSO₂-aryl, —NHSO₂-heteroaryl,—NHSO₂-heterocycloalkyl, —CH₂NH₂, —CH₂SO₂CH₃, -aryl, -arylalkyl,-heteroaryl, -heteroarylalkyl, -heterocycloalkyl, —C₃-C₁₂-cycloalkyl,polyalkoxyalkyl, polyalkoxy, -methoxymethoxy, -methoxyethoxy, —SH,—S—C₁-C₁₂-alkyl, —S—C₂-C₁₂-alkenyl, —S—C₂-C₁₂-alkenyl,—S—C₃-C₁₂-cycloalkyl, —S-aryl, —S-heteroaryl, —S-heterocycloalkyl, ormethylthiomethyl.

It is understood that the aryls, heteroaryls, alkyls, and the like canbe substituted.

The term “cancer” includes, but is not limited to, the followingcancers: epidermoid Oral: buccal cavity, lip, tongue, mouth, pharynx;Cardiac: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma,liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma, and teratoma: Lung:bronchogenic carcinoma (squamous cell or epidermoid, undifferentiatedsmall cell, undifferentiated large cell, adenocarcinoma), alveolar(bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma,chondromatous hamartoma, mesothelioma; Gastrointestinal: esophagus(squamous cell carcinoma, larynx, adenocarcinoma, leiomyosarcoma,lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas(ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoidtumors, vipoma), small bowel or small intestines (adenocarcinoma,lymphoma, carcinoid tumors, Karposi's sarcoma, leiomyoma, hemangioma,lipoma, neurofibroma, fibroma), large bowel or large intestines(adenocarcinoma, tubular adenoma, vinous adenoma, hamartoma, leiomyoma),colon, colon-rectum, colorectal, rectum; Genitourinary tract: kidney(adenocarcinoma, Wilm's tumor (nephroblastoma), lymphoma, leukemia),bladder and urethra (squamous cell carcinoma, transitional cellcarcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis(seminoma, teratoma, embryonal carcinoma, teratocarcinoma,choriocarcinoma, sarcoma, interstitial cell carcinoma, fibroma,fibroadenoma, adenomatoid tumors, lipoma); Liver: hepatoma(hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma,angiosarcoma, hepatocellular adenoma, hemangioma, biliary passages;Bone: osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibroushistiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma(reticulum cell sarcoma), multiple myeloma, malignant giant cell tumorchordoma, osteochronfroma (osteocartilaginous exostoses), benignchondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma andgiant cell tumors; Nervous system: skull (osteoma, hemangioma,granuloma, xanthoma, osteitis deformans), meninges (meningioma,meningiosarcoma, gliomatosis), brain (astrocytoma, medulloblastoma,glioma, ependymoma, germinoma (pinealoma), glioblastoma multiform,oligodendroglioma, schwannoma, retinoblastoma, congenital tumors),spinal cord neurofibroma, meningioma, glioma, sarcoma); Gynecological:uterus (endometrial carcinoma), cervix (cervical carcinoma, pre-tumorcervical dysplasia), ovaries (ovarian carcinoma (serouscystadenocarcinoma, mucinous cystadenocarcinoma, unclassifiedcarcinoma), granulosa-thecal cell tumors, Sertoli-Leydig cell tumors,dysgerminoma, malignant teratoma), vulva (squamous cell carcinoma,intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma),vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma(embryonal rhabdomyosarcoma), fallopian tubes (carcinoma), breast;Hematologic: blood (myeloid leukemia (acute and chronic), acutelymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferativediseases, multiple myeloma, myelodysplastic syndrome). Hodgkin'sdisease, non-Hodgkin's lymphoma (malignant lymphoma) hairy cell;lymphoid disorders; Skin: malignant melanoma, basal cell carcinoma,squamous cell carcinoma, Karposi's sarcoma, keratoacanthoma, molesdysplastic nevi, lipoma, angioma, dermatofibroma, keloids, psoriasis.Thyroid gland: papillary thyroid carcinoma, follicular thyroidcarcinoma; medullary thyroid carcinoma, undifferentiated thyroid cancer,multiple endocrine neoplasia type 2A, multiple endocrine neoplasia type2B, familial medullary thyroid cancer, pheochromocytoma, paraganglioma:and Adrenal glands: neuroblastoma. Thus, the term “cancerous cell” asprovided herein, includes a cell afflicted by any one of theabove-identified conditions.

The term “EGFR” herein refers to epidermal growth factor receptorkinase.

The term “HER” or “Her”, herein refers to human epidermal growth factorreceptor kinase.

The term “subject” as used herein refers to a mammal. A subjecttherefore refers to, for example, dogs, cats, horses, cows, pigs, guineapigs, and the like. Preferably the subject is a human. When the subjectis a human, the subject may be referred to herein as a patient.

“Treat”, “treating” and “treatment” refer to a method of alleviating orabating a disease and/or its attendant symptoms.

As used herein, “preventing” or “prevent” describes reducing oreliminating the onset of the symptoms or complications of the disease,condition or disorder.

As used herein, the term “allosteric site” refers to a site on EGFRother than the ATP binding site, such as that characterized in a crystalstructure of EGFR. An “allosteric site” can be a site that is close tothe ATP binding site, such as that characterized in a crystal structureof EGFR. For example, one allosteric site includes one or more of thefollowing amino acid residues of EGFR: Lys745, Leu788, Ala 743, Cys755,Leu777, Phe856, Asp855, Met766, Ile759, Glu762, and/or Ala763.

As used herein, the term “allosteric EGFR inhibitor” refers to acompound that inhibits EGFR activity through binding to one or moreallosteric sites on EGFR.

As used herein, the term “agent that prevents EGFR dimer formation”refers to an agent that prevents dimer formation in which the C-lobe ofthe “activator” subunit impinges on the N-lobe of the “receiver”subunit. Examples of agents that prevent EGFR dimer formation include,but are not limited to, cetuximab, cobimetinib, trastuzumab,panitumumab, and Mig6.

As used herein the term “GDC0973” or “Cobimetinib” refers to a compoundhaving the chemical structure:

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts of the compounds formed by the process of the presentapplication which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art. For example, S. M. Berge, etal., describes pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences, 66: 1-19 (1977). The salts can be prepared insitu during the final isolation and purification of the compounds of theapplication, or separately by reacting the free base function with asuitable organic acid.

Examples of pharmaceutically acceptable include, but are not limited to,nontoxic acid addition salts are salts of an amino group formed withinorganic acids such as hydrochloric acid, hydrobromic acid, phosphoricacid, sulfuric acid and perchloric acid or with organic acids such asacetic acid, maleic acid, tartaric acid, citric acid, succinic acid ormalonic acid or by using other methods used in the art such as ionexchange. Other pharmaceutically acceptable salts include, but are notlimited to, adipate, alginate, ascorbate, aspartate, benzenesulfonate,benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate,citrate, cyclopentanepropionate, digluconate, dodecylsulfate,ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate,gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and thelike. Representative alkali or alkaline earth metal salts includesodium, lithium, potassium, calcium, magnesium, and the like. Furtherpharmaceutically acceptable salts include, when appropriate, nontoxicammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, alkyl having from 1 to 6 carbon atoms, sulfonate and arylsulfonate.

As used herein, the term “pharmaceutically acceptable ester” refers toesters of the compounds formed by the process of the present applicationwhich hydrolyze in vivo and include those that break down readily in thehuman body to leave the parent compound or a salt thereof. Suitableester groups include, for example, those derived from pharmaceuticallyacceptable aliphatic carboxylic acids, particularly alkanoic, alkenoic,cycloalkanoic and alkanedioic acids, in which each alkyl or alkenylmoiety advantageously has not more than 6 carbon atoms. Examples ofparticular esters include, but are not limited to, formates, acetates,propionates, butyrates, acrylates and ethylsuccinates.

The term “pharmaceutically acceptable prodrugs” as used herein refers tothose prodrugs of the compounds formed by the process of the presentapplication which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswith undue toxicity, irritation, allergic response, and the like,commensurate with a reasonable benefit/risk ratio, and effective fortheir intended use, as well as the zwitterionic forms, where possible,of the compounds of the present application. “Prodrug”, as used hereinmeans a compound which is convertible in vivo by metabolic means (e.g.,by hydrolysis) to afford any compound delineated by the formulae of theinstant application. Various forms of prodrugs are known in the art, forexample, as discussed in Bundgaard, (ed.), Design of Prodrugs, Elsevier(1985); Widder, et al., (ed.), Methods in Enzymology, vol. 4, AcademicPress (1985); Krogsgaard-Larsen, et al., (ed). Design and Application ofProdrugs, Textbook of Drug Design and Development, Chapter 5, 113-191(1991); Bundgaard, et at, Journal of Drug Deliver Reviews, 8:1-38(1992);Bundgaard, J. of Pharmaceutical Sciences, 77:285 et seq. (1988); Higuchiand Stella (eds.) Prodrugs as Novel Drug Delivery Systems, AmericanChemical Society (1975); and Bernard Testa & Joachim Mayer, “HydrolysisIn Drug And Prodrug Metabolism: Chemistry, Biochemistry And Enzymology,”John Wiley and Sons, Ltd. (2002).

This application also encompasses pharmaceutical compositionscontaining, and methods of treating disorders through administering,pharmaceutically acceptable prodrugs of compounds of the application.For example, compounds of the application having free amino, amido,hydroxy or carboxylic groups can be converted into prodrugs. Prodrugsinclude compounds wherein an amino acid residue, or a polypeptide chainof two or more (e.g., two, three or four) amino acid residues iscovalently joined through an amide or ester bond to a free amino,hydroxy or carboxylic acid group of compounds of the application. Theamino acid residues include but am not limited to the 20 naturallyoccurring amino acids commonly designated by three letter symbols andalso includes 4-hydroxyproline, hydroxylysine, demosine, isodemosine,3-methylhistidine, norvalin, beta-alanine, gamma-aminobutyric acid,citrulline, homocysteine, homoserine, ornithine and methionine sulfone.Additional types of prodrugs are also encompassed. For instance, freecarboxyl groups can be derivatized as amides or alkyl esters. Freehydroxy groups may be derivatized using groups including but not limitedto hemisuccinates, phosphate esters, dimethylaminoacetates, andphosphoryloxymethyloxy carbonyls, as outlined in Advanced Drug DeliveryReviews, 1996, 19, 1 15. Carbamate prodrugs of hydroxy and amino groupsare also included, as are carbonate prodrugs, sulfonate esters andsulfate esters of hydroxy groups. Derivatization of hydroxy groups as(acyloxy)methyl and (acyloxy)ethyl ethers wherein the acyl group may bean alkyl ester, optionally substituted with groups including but notlimited to ether, amine and carboxylic acid functionalities, or wherethe acyl group is an amino acid ester as described above, are alsoencompassed. Prodrugs of this type are described in J. Med Chem. 1996,39, 10. Free amines can also be derivatized as amides, sulfonamides orphosphonamides. All of these prodrug moieties may incorporate groupsincluding but not limited to ether, amine and carboxylic acidfunctionalities

Combinations of substituents and variables envisioned by thisapplication are only those that result in the formation of stablecompounds. The term “stable”, as used herein, refers to compounds whichpossess stability sufficient to allow manufacture and which maintainsthe integrity of the compound for a sufficient period of time to beuseful for the purposes detailed herein (e.g., therapeutic orprophylactic administration to a subject).

In addition, some of the compounds of this application have one or moredouble bonds, or one or more asymmetric centers. Such compounds canoccur as racemates, racemic mixtures, single enantiomers, individualdiastereomers, diastereomeric mixtures, and cis- or trans- or E- orZ-double isomeric forms, and other stereoisomeric forms that may bedefined, in terms of absolute stereochemistry, as (R)- or (S)-, or as(D)- or (L)- for amino acids. All such isomeric forms of these compoundsare expressly included in the present application.

“Isomerism” means compounds that have identical molecular formulae butdiffer in the sequence of bonding of their atoms or in the arrangementof their atoms in space. Isomers that differ in the arrangement of theiratoms in space are termed “stereoisomers”. Stereoisomers that are notmirror images of one another are termed “diastereoisomers”, andstereoisomers that are non-superimposable mirror images of each otherare termed “enantiomers” or sometimes optical isomers. A mixturecontaining equal amounts of individual enantiomeric forms of oppositechirality is termed a “racemic mixture”.

A carbon atom bonded to four non-identical substituents is termed a“chiral center”. “Chiral isomer” means a compound with at least onechiral center. Compounds with more than one chiral center may existeither as an individual diastereomer or as a mixture of diastereomers,termed “diastereomeric mixture”. When one chiral center is present, astereoisomer may be characterized by the absolute configuration (R or S)of that chiral center. Absolute configuration refers to the arrangementin space of the substituents attached to the chiral center. Thesubstituents attached to the chiral center under consideration areranked in accordance with the Sequence Rule of Cahn, Ingold and Prelog.(Cahn et al., Angew. Chem. Inter. Edit. 1966, 5, 385; errata 511; Calmet al., Angew. Chem. 1966, 78, 413; Calm and Ingold, J. Chem. Soc. 1951(London), 612; Cahn et al., Experientia 1956, 12, 81: Cahn, J. Chem.Educ. 1964, 41, 116).

“Geometric isomer” means the diastereomers that owe their existence tohindered rotation about double bonds. These configurations aredifferentiated in their names by the prefixes cis and trans, or Z and E,which indicate that the groups are on the same or opposite side of thedouble bond in the molecule according to the Cahn-Ingold-Prelog rules.

Furthermore, the structures and other compounds discussed in thisapplication include all atropic isomers thereof. “Atropic isomers” are atype of stereoisomer in which the atoms of two isomers are arrangeddifferently in space. Atropic isomers owe their existence to arestricted rotation caused by hindrance of rotation of large groupsabout a central bond. Such atropic isomers typically exist as a mixture,however as a result of recent advances in chromatography techniques: ithas been possible to separate mixtures of two atropic isomers in selectcases.

“Tautomer” is one of two or more structural isomers that exist inequilibrium and is readily converted from one isomeric form to another.This conversion results in the formal migration of a hydrogen atomaccompanied by a switch of adjacent conjugated double bonds. Tautomersexist as a mixture of a tautomeric set in solution. In solid form,usually one tautomer predominates. In solutions where tautomerization ispossible, a chemical equilibrium of the tautomers will be reached. Theexact ratio of the tautomers depends on several factors, includingtemperature, solvent and pH. The concept of tautomers that aminterconvertable by tautomerizations is called tautomerism.

Of the various types of tautomerism that are possible, two are commonlyobserved. In keto-enol tautomerism a simultaneous shift of electrons anda hydrogen atom occurs. Ring-chain tautomerism arises as a result of thealdehyde group (—CHO) in a sugar chain molecule reacting with one of thehydroxy groups (—OH) in the same molecule to give it a cyclic(ring-shaped) form as exhibited by glucose. Common tautomeric pairs are:ketone-enol, amide-nitrile, lactam-lactim, amide-imidic acid tautomerismin heterocyclic rings (e.g., in nucleobases such as guanine, thymine andcytosine), amine-enamine and enamine-enamine.

The compounds of this application may also be represented in multipletautomeric forms, in such instances, the application expressly includesall tautomeric forms of the compounds described herein (e.g., alkylationof a ring system may result in alkylation at multiple sites, theapplication expressly includes all such reaction products). When thecompounds described herein contain olefinic double bonds or othercenters of geometric asymmetry, and unless specified otherwise, it isintended that the compounds include both E and Z geometric isomers.Likewise, all tautomeric forms are also intended to be included. Theconfiguration of any carbon-carbon double bond appearing herein isselected for convenience only and is not intended to designate aparticular configuration unless the text so states; thus a carbon-carbondouble bond depicted arbitrarily herein as tram may be cis, tram, or amixture of the two in any proportion. All such isomeric forms of suchcompounds are expressly included in the present application.

In the present specification, the structural formula of the compoundrepresents a certain isomer for convenience in some cases, but thepresent application includes all isomers, such as geometrical isomers,optical isomers based on an asymmetrical carbon, stereoisomers,tautomers, and the like.

Furthermore, so-called metabolite which is produced by degradation ofthe present compound in vivo is included in the scope of the presentapplication.

The term “crystal polymorphs”, “polymorphs” or “crystal forms” meanscrystal structures in which a compound (or a salt solvate, orstereoisomer thereof) can crystallize in different crystal packingarrangements, all of which have the same elemental composition.Different crystal forms usually have different X-ray diffractionpatterns, infrared spectral, melting points, density hardness, crystalshape, optical and electrical properties, stability and solubility.Recrystallization solvent, rate of crystallization, storage temperature,and other factors may cause one crystal form to dominate. Crystalpolymorphs of the compounds can be prepared by crystallization underdifferent conditions.

Additionally, the compounds of the present application, for example, thesalts of the compounds, can exist in either hydrated or unhydrated (theanhydrous) form or as solvates with other solvent molecules.Non-limiting examples of hydrates include monohydrates, dihydrates, etc.Non-limiting examples of solvates include ethanol solvates, acetonesolvates, etc.

“Solvate” means solvent addition forms that contain eitherstoichiometric or non stoichiometric amounts of solvent. Some compoundshave a tendency to trap a fixed molar ratio of solvent molecules in thecrystalline solid state, thus forming a solvate. If the solvent is waterthe solvate formed is a hydrate; and if the solvent is alcohol, thesolvate formed is an alcoholate. Hydrates are formed by the combinationof one or more molecules of water with one molecule of the substance inwhich the water retains its molecular state as H₂O.

Method of Synthesizing the Compounds

The compounds of the present application may be made by a variety ofmethods, including standard chemistry. The synthetic processes of theapplication can tolerate a wide variety of functional groups, thereforevarious substituted starting materials can be used. The processesgenerally provide the desired final compound at or near the end of theoverall process, although it may be desirable in certain instances tofurther convert the compound to a pharmaceutically acceptable salt,ester or prodrug thereof. Suitable synthetic routes are depicted in theschemes below.

Compounds of the present application can be prepared in a variety ofways using commercially available starting materials, compounds known inthe literature, or from readily prepared intermediates, by employingstandard synthetic methods and procedures either known to those skilledin the art, or which will be apparent to the skilled artisan in light ofthe teachings herein. Standard synthetic methods and procedures for thepreparation of organic molecules and functional group transformationsand manipulations can be obtained from the relevant scientificliterature or from standard textbooks in the field. Although not limitedto any one or several sources, classic texts such as Smith, M. B.,March, J., March's Advanced Organic Chemistry: Reactions. Mechanisms,and Structure, 5^(th) edition, John Wiley & Sons: New York, 2001; andGreene, T. W., Wuts, P. G. M., Protective Groups in Organic Synthesis,3^(rd) edition, John Wiley & Sons: New York, 1999, incorporated byreference herein, am useful and recognized reference textbooks oforganic synthesis known to those in the art. The following descriptionsof synthetic methods are designed to illustrate, but not to limit,general procedures for the preparation of compounds of the presentapplication.

The compounds of disclosed herein may be prepared by methods known inthe art of organic synthesis as set forth in part by the followingsynthetic schemes. In the schemes described below, it is well understoodthat protecting groups for sensitive or reactive groups are employedwhere necessary in accordance with general principles or chemistry.Protecting groups are manipulated according to standard methods oforganic synthesis (T. W. Greene and P. G. M. Wuts, “Protective Groups inOrganic Synthesis”, Third edition, Wiley, New York 1999). These groupsare removed at a convenient stage of the compound synthesis usingmethods that are readily apparent to those skilled in the art. Theselection processes, as well as the reaction conditions and order oftheir execution, shall be consistent with the preparation of compoundsof disclosed herein.

Those skilled in the art will recognize if a stereocenter exists in thecompounds of disclosed herein. Accordingly, the present applicationincludes both possible stereoisomers (unless specified in the synthesis)and includes not only racemic compounds but the individual enantiomersand/or diastereomers as well. When a compound is desired as a singleenantiomer or diastereomer, it may be obtained by stereospecificsynthesis or by resolution of the final product or any convenientintermediate. Resolution of the final product, an intermediate, or astarting material may be affected by any suitable method known in theart. See, for example, “Stereochemistry of Organic Compounds” by E. L.Eliel, S. H. Wilen, and L. N. Mander (Wiley-Interscience, 1994).

All the abbreviations used in this application are found in “ProtectiveGroups in Organic Synthesis” by John Wiley & Sons, Inc, or the MERCKINDEX by MERCK R. Co., Inc, or other chemistry books or chemicalscatalogs by chemicals vendor such as Aldrich, or according to usage knowin the art.

The compounds of the present application can be prepared in a number ofways well known to those skilled in the art of organic synthesis. By wayof example, compounds of the present application can be synthesizedusing the methods described below, together with synthetic methods knownin the art of synthetic organic chemistry, or variations thereon asappreciated by those skilled in the art. Preferred methods include butare not limited to those methods described below. Compounds of thepresent application can be synthesized by following the steps outlinedin General Schemes 1-3 which comprise different sequences of assemblingintermediates and compounds of the application. Starting materials areeither commercially available or made by known procedures in thereported literature or as illustrated.

Compounds of the application (e.g., a compound of Formula I) may beprepared according to General Schemes 1, 2, and 3a-e under appropriateconditions, such as those exemplified in the Examples.

A mixture of enantiomers, diastereomers, and/or cis/trans isomersresulting from the processes described above can be separated into theirsingle components by chiral salt technique, chromatography using normalphase, or reverse phase or chiral column, depending on the nature of theseparation.

It should be understood that in the description and formulae shownabove, the various groups and other variables are as defined herein,except where otherwise indicated. Furthermore, for synthetic purposes,the compounds of General Schemes are mere representatives with electedradicals to illustrate the general synthetic methodology of thecompounds of disclosed herein.

A compound of the application can be prepared as a pharmaceuticallyacceptable acid addition salt by reacting the free base form of thecompound with a pharmaceutically acceptable inorganic or organic acid.Alternatively, a pharmaceutically acceptable base addition salt of acompound of the application can be prepared by reacting the free acidform of the compound with a pharmaceutically acceptable inorganic ororganic base. Alternatively, the salt forms of the compounds of theapplication can be prepared using salts of the starting materials orintermediates.

The free acid or free base forms of the compounds of the application canbe prepared from the corresponding base addition salt or acid additionsalt from, respectively. For example a compound of the application in anacid addition salt form can be converted to the corresponding free baseby treating with a suitable base (e.g., ammonium hydroxide solution,sodium hydroxide, and the like). A compound of the application in a baseaddition salt form can be converted to the corresponding free acid bytreating with a suitable acid (e.g., hydrochloric acid, etc.).

Prodrugs of the compounds of the application can be prepared by methodsknown to those of ordinary skill in the art (e.g., for further detailssee Saulnier et al., (1994), Bioorganic and Medicinal Chemistry Letters,Vol. 4, p. 1985). For example, appropriate prodrugs can be prepared byreacting a non-derivatized compound of the application with a suitablecarbamylating agent (e.g., 1,1-acyloxyalkylcarbanochloridate,para-nitrophenyl carbonate, or the like).

Protected derivatives of the compounds of the application can be made bymeans known to those of ordinary skill in the art. A detaileddescription of techniques applicable to the creation of protectinggroups and their removal can be found in T. W. Greene, “ProtectingGroups in Organic Chemistry”, 3rd edition, John Wiley and Sons, Inc.,1999.

Compounds of the present application can be conveniently prepared, orformed during the process of the application, as solvates (e.g.,hydrates). Hydrates of compounds of the present application can beconveniently prepared by recrystallization from an aqueous/organicsolvent mixture, using organic solvents such as dioxin, tetrahydrofuranor methanol.

Acids and bases useful in the methods herein are known in the art. Acidcatalysts are any acidic chemical, which can be inorganic (e.g.,hydrochloric, sulfuric, nitric acids, aluminum trichloride) or organic(e.g., camphorsulfonic acid, p-toluenesulfonic acid, acetic acid,ytterbium triflate) in nature. Acids are useful in either catalytic orstoichiometric amounts to facilitate chemical reactions. Bases are anybasic chemical, which can be inorganic (e.g., sodium bicarbonate,potassium hydroxide) or organic (e.g., triethylamine, pyridine) innature. Bases are useful in either catalytic or stoichiometric amountsto facilitate chemical reactions.

Optical isomers may be prepared from their respective optically activeprecursors by the procedures described herein, or by resolving theracemic mixtures. The resolution can be carried out in the presence of aresolving agent, by chromatography or by repeated crystallization or bysome combination of these techniques which are known to those skilled inthe art. Further details regarding resolutions can be found in Jacques,et al., Enantiomers, Racemates, and Resolutions (John Wiley & Sons,1981).

The synthesized compounds can be separated from a reaction mixture andfurther purified by a method such as column chromatography, highpressure liquid chromatography, or recrystallization. As can beappreciated by the skilled artisan, further methods of synthesizing thecompounds of the formulae herein will be evident to those of ordinaryskill in the art. Additionally, the various synthetic steps may beperformed in an alternate sequence or order to give the desiredcompounds. In addition, the solvents, temperatures, reaction durations,etc. delineated herein are for purposes of illustration only and one ofordinary skill in the art will recognize that variation of the reactionconditions can produce the desired bridged macrocyclic products of thepresent application. Synthetic chemistry transformations and protectinggroup methodologies (protection and deprotection) useful in synthesizingthe compounds described herein am known in the art and include, forexample, those such as described in R. Larock, Comprehensive OrganicTransformations, VCH Publishers (1989); T. W. Greene and P. G. M. Wuts,Protective Groups in Organic Synthesis, 2d. Ed., John Wiley and Sons(1991); L. Fieser and M. Fieser, Fieser and Fieser's Reagents forOrganic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed.,Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons(1995), and subsequent editions thereof.

The compounds of this application may be modified by appending variousfunctionalities via any synthetic means delineated herein to enhanceselective biological properties. Such modifications are known in the artand include those which increase biological penetration into a givenbiological system (e.g., blood, lymphatic system, central nervoussystem), increase oral availability, increase solubility to allowadministration by injection, alter metabolism and alter rate ofexcretion.

Biological Assays Biochemical Assays

EGFR biochemical assays are carried out using a homogeneoustime-resolved fluorescence (HTRF) assay. The reaction mixtures containbiotin-Lck-peptide substrate, wild type, or mutant EGFR enzyme inreaction buffer. Enzyme concentrations are adjusted to accommodatevarying kinase activity and ATP concentrations. Compounds of the presentapplication are diluted into the assay mixture and IC₅₀ values aredetermined using 12-point inhibition curves.

Phospho-EGFR Target Modulation Assays and ELISA

Cells are lysed with lysis buffer containing protease and phosphataseinhibitors and the plates are shaken. An aliquot from each well is thentransferred to prepared ELISA plates for analysis. Once harvested andplated, the cells are pre-treated with media with or without EGF. Thecompounds of the present application are then added and IC₅₀ values aredetermined using an EGFR biochemical assay described above.

Solid high-binding ELISA plates are coated with goat anti-EGFR captureantibody. Plates are then blocked with BSA in a buffer, and then washed.Aliquots of lysed cell are added to each well of the ELISA plate and theplate is incubated. An anti-phospho-EGFR is then added and is followedby further incubation. After washing, anti-rabbit-HRP is added and theplate is again incubated. Chemiluminescent detection is carried out withSuperSignal ELISA Pico substrate. Signal is read on EnVision platereader using built-in UltraLUM setting.

Western Blotting

Cell lysates are equalized to protein content and loaded onto a gel withrunning buffer. Membranes are probed with primary antibodies and arethen washed. HRP-conjugated secondary antibodies are added and afterwashing. HRP is detected using a HRP substrate reagent and recorded withan imager.

Cell Proliferation Assays

Cell lines are plated in media. The compounds of the present applicationare then serially diluted and transferred to the cells. Cell viabilityis measured via a luminescent readout. Data is analyzed by non-linearregression curve-fitting.

Methods of the Application

In another aspect, the application provides a method of inhibiting akinase, comprising contacting the kinase with a compound disclosedherein, or a pharmaceutically acceptable salt, hydrate, solvate, orstereoisomer thereof. In some embodiments, the kinase comprises amutated cysteine residue. In further embodiments, the mutated cysteineresidue is located in or near the position equivalent to Cys 797 inEGFR, including such position in Jak3, Blk, Bmx, Btk, HER2 (ErbB2), HER4(ErbB4), Itk, Tec, and Txk. In some embodiments, the kinase is EGFR. Insome embodiments, the kinase is a Her-kinase. In other embodiments, themethod further comprises a second agent that prevents kinase dimerformation. In some embodiments, the second agent that prevents kinasedimer formation is an antibody. In further embodiments, the second agentprevents EGFR dimer formation. In further embodiments, the second agentthat prevents EGFR dimer formation is cetuximab, trastuzumab, orpanitumumab. In further embodiments, the second agent that prevents EGFRdimer formation is cetuximab.

In another aspect, the application provides a method of inhibiting akinase, the method comprising administering to a subject in need thereofan effective amount of a compound disclosed herein, or apharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof. In some embodiments, the kinase is EGFR. In some embodiments,the kinase is a Her-kinase. In other embodiments, the method furthercomprises administering a second agent that prevents dimer formation ofthe kinase. In some embodiments, the second agent that prevents kinasedimer formation is an antibody. In further embodiments, the second agentprevents EGFR dimer formation. In further embodiments, the second agentthat prevents EGFR dimer formation is cetuximab, trastuzumab, orpanitumumab. In further embodiments, the second agent that prevents EGFRdimer formation is cetuximab.

In still another aspect, the application provides a method of inhibitingEGFR, the method comprising administering to a subject in need thereofan effective amount of a compound disclosed herein, or apharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof. In some embodiments, the method further comprises administeringa second agent that prevents EGFR dimer formation. In some embodiments,the second agent that prevents EGFR dimer formation is an antibody. Infurther embodiments, the second agent that prevents EGFR dimer formationis cetuximab, trastuzumab, or panitumumab. In further embodiments, thesecond agent that prevents EGFR dimer formation is cetuximab.

Another aspect of the application provides a method of treating orpreventing a disease, the method comprising administering to a subjectin need thereof an effective amount of a compound disclosed herein, or apharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof. In some embodiments, the disease is mediated by a kinase. Infurther embodiments, the kinase comprises a mutated cysteine residue. Infurther embodiments, the mutated cysteine residue is located in or nearthe position equivalent to Cys 797 in EGFR, including such positions inJak3, Blk, Bmx, Btk. HER2 (ErbB2), HER4 (ErbB4), Itk, Tec, and Txk. Insome embodiments, the method further comprises administering a secondagent that prevents dimer formation of the kinase. In some embodiments,the second agent that prevents kinase dimer formation is an antibody. Infurther embodiments, the second agent prevents EGFR dimer formation. Infurther embodiments, the second agent that prevents EGFR dimer formationis cetuximab, trastuzumab, or panitumumab. In further embodiments, thesecond agent that prevents EGFR dimer formation is cetuximab.

In some embodiments, the disease is mediated by EGFR (e.g., EGFR plays arole in the initiation or development of the disease). In furtherembodiments, the EGFR is a Her-kinase. In further embodiments, theHer-kinase is HER1, HER2, or HER4. In some embodiments, the EGFRcomprises one or more mutations, as described herein.

In certain embodiments, the disease is cancer or a proliferationdisease.

In further embodiments, the disease is lung cancer, colon cancer, breastcancer, prostate cancer, liver cancer, pancreas cancer, brain cancer,kidney cancer, ovarian cancer, stomach cancer, skin cancer, bone cancer,gastric cancer, breast cancer, pancreatic cancer, glioma, glioblastoma,hepatocellular carcinoma, papillary renal carcinoma, head and necksquamous cell carcinoma, leukemias, lymphomas, myelomas, or solidtumors.

In other embodiments, the disease is inflammation, arthritis, rheumatoidarthritis, spondyiarthropathies, gouty arthritis, osteoarthritis,juvenile arthritis, and other arthritic conditions, systemic lupuserthematosus (SLE), skin-related conditions, psoriasis, eczema, burns,dermatitis, neuroinflammation, allergy, pain, neuropathic pain, fever,pulmonary disorders, lung inflammation, adult respiratory distresssyndrome, pulmonary sarcoisosis, asthma, silicosis, chronic pulmonaryinflammatory disease, and chronic obstructive pulmonary disease (COPD),cardiovascular disease, arteriosclerosis, myocardial infarction(including post-myocardial infarction indications), thrombosis,congestive heart failure, cardiac reperfusion injury, as well ascomplications associated with hypertension and/or heart failure such asvascular organ damage, restenosis, cardiomyopathy, stroke includingischemic and hemorrhagic stroke, reperfusion injury, renal reperfusioninjury, ischemia including stroke and brain ischemia, and ischemiaresulting from cardiac/coronary bypass, neurodegenerative disorders,liver disease and nephritis, gastrointestinal conditions, inflammatorybowel disease, Crohn's disease, gastritis, irritable bowel syndrome,ulcerative colitis, ulcerative diseases, gastric ulcers, viral andbacterial infections, sepsis, septic shock, gram negative sepsis,malaria, meningitis. HIV infection, opportunistic infections, cachexiasecondary to infection or malignancy, cachexia secondary to acquiredimmune deficiency syndrome (AIDS), AIDS, ARC (AIDS related complex),pneumonia, herpes virus, myalgias due to infection, influenza,autoimmune disease, graft vs. host reaction and allograft rejections,treatment of bone resorption diseases, osteoporosis, multiple sclerosis,cancer, leukemia, lymphoma, colorectal cancer, brain cancer, bonecancer, epithelial call-derived neoplasia (epithelial carcinoma), basalcell carcinoma, adenocarcinoma, gastrointestinal cancer, lip cancer,mouth cancer, esophageal cancer, small bowel cancer, stomach cancer,colon cancer, liver cancer, bladder cancer, pancreas cancer, ovariancancer, cervical cancer, lung cancer, breast cancer, skin cancer,squamus cell and/or basal cell cancers, prostate cancer, renal cellcarcinoma, and other known cancers that affect epithelial cellsthroughout the body, chronic myelogenous leukemia (CML), acute myeloidleukemia (AML) and acute promyelocytic leukemia (APL), angiogenesisincluding neoplasia, metastasis, central nervous system disorders,central nervous system disorders having an inflammatory or apoptoticcomponent, Alzheimer's disease. Parkinson's disease. Huntington'sdisease, amyotrophic lateral sclerosis, spinal cord injury, andperipheral neuropathy, or B-Cell Lymphoma.

In further embodiments, the disease is inflammation, arthritis,rheumatoid arthritis, spondylarthropathies, gouty arthritis,osteoarthritis, juvenile arthritis, and other arthritic conditions,systemic lupus erthematosus (SLE), skin-related conditions, psoriasis,eczema, dermatitis, pain, pulmonary disorders, lung inflammation, adultrespiratory distress syndrome, pulmonary sarcoisosis, asthma, chronicpulmonary inflammatory disease, and chronic obstructive pulmonarydisease (COPD), cardiovascular disease, arteriosclerosis, myocardialinfarction (including post-myocardial infarction indications),congestive heart failure, cardiac reperfusion injury, inflammatory boweldisease. Crohn's disease, gastritis, irritable bowel syndrome, leukemiaor lymphoma.

Another aspect of the application provides a method of treating a kinasemediated disorder, the method comprising administering to a subject inneed thereof an effective amount of a compound disclosed herein, or apharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof. In other embodiments, the compound is an inhibitor of HER1,HER2, or HER4. In other embodiments, the subject is administered anadditional therapeutic agent. In other embodiments, the compound and theadditional therapeutic agent are administered simultaneously orsequentially.

In another aspect, the application provides a method of treating akinase mediated disorder, the method comprising administering to asubject in need thereof an effective amount of a compound disclosedherein, or a pharmaceutically acceptable salt, hydrate, solvate, orstereoisomer thereof, and a second agent that prevents EGFR dimerformation. In other embodiments, the compound is an inhibitor of HER1,HER2, or HER4. In other embodiments, the subject is administered anadditional therapeutic agent. In other embodiments, the compound, thesecond agent that prevents EGFR dimer formation, and the additionaltherapeutic agent are administered simultaneously or sequentially. Insome embodiments, the second agent that prevents EGFR dimer formation isan antibody. In further embodiments, the second agent that prevents EGFRdimer formation is cetuximab, trastuzumab, or panitumumab. In furtherembodiments, the second agent that prevents EGFR dimer formation iscetuximab.

In other embodiments, the disease is cancer. In further embodiments, thecancer is lung cancer, colon cancer, breast cancer, prostate cancer,liver cancer, pancreas cancer, brain cancer, kidney cancer, ovariancancer, stomach cancer, skin cancer, bone cancer, gastric cancer, breastcancer, pancreatic cancer, glioma, glioblastoma, hepatocellularcarcinoma, papillary renal carcinoma, head and neck squamous cellcarcinoma, leukemias, lymphomas, myelomas, or solid tumors.

In another aspect, the application provides a method of treating orpreventing cancer, wherein the cancer cell comprise activated EGFR,comprising administering to a subject in need thereof an effectiveamount of a compound disclosed herein, or a pharmaceutically acceptablesalt, hydrate, solvate, or stereoisomer thereof.

In another aspect, the application provides a method of treating orpreventing cancer, wherein the cancer cell comprise activated EGFR,comprising administering to a subject in need thereof an effectiveamount of a compound disclosed herein, or a pharmaceutically acceptablesalt, hydrate, solvate, or stereoisomer thereof and a second agent thatprevents EGFR dimer formation. In some embodiments, the second agentthat prevents EGFR dimer formation is an antibody. In furtherembodiments, the second agent that prevents EGFR dimer formation iscetuximab, trastuzumab, or panitumumab. In further embodiments, thesecond agent that prevents EGFR dimer formation is cetuximab.

In certain embodiments, the EGFR activation is selected from mutation ofEGFR, amplification of EGFR, expression of EGFR, and ligand mediatedactivation of EGFR.

Another aspect of the application provides a method of treating orpreventing cancer in a subject, wherein the subject is identified asbeing in need of EGFR inhibition for the treatment of cancer, comprisingadministering to the subject an effective amount of a compound disclosedherein, or a pharmaceutically acceptable salt, hydrate, solvate, orstereoisomer thereof.

In another aspect, the application provides a method of treating orpreventing cancer in a subject, wherein the subject is identified asbeing in need of EGFR inhibition for the treatment of cancer, comprisingadministering to the subject an effective amount of a compound disclosedherein, or a pharmaceutically acceptable salt, hydrate, solvate, orstereoisomer thereof, and optionally a second agent that prevents EGFRdimer formation. In some embodiments, the second agent that preventsEGFR dimer formation is an antibody. In further embodiments, the secondagent that prevents EGFR dimer formation is cetuximab, trastuzumab, orpanitumumab. In further embodiments, the second agent that prevents EGFRdimer formation is cetuximab.

In certain embodiments, the subject identified as being in need of EGFRinhibition is resistant to a known EGFR inhibitor, including but notlimited to, gefitinib, erlotinib, afatinib. AZD9291, CO-1686, or WZ4002.In certain embodiments, a diagnostic test is performed to determine ifthe subject has an activating mutation in EGFR. In certain embodiments,a diagnostic test is performed to determine if the subject has an EGFRharboring an activating and a drug resistance mutation, such as thosedescribed herein. Activating mutations comprise without limitationL858R, G719S, G719C, G719A, L718Q, L861Q, a deletion in exon 19 and/oran insertion in exon 20. Drug resistant EGFR mutants can have withoutlimitation a drug resistance mutation comprising T790M, T854A, L718Q,C797S, or D761Y. The diagnostic test can comprise sequencing,pyrosequencing, PCR, RT-PCR, or similar analysis techniques known tothose of skill in the art that can detect nucleotide sequences.

In another aspect, the application provides a method of treating orpreventing cancer, wherein the cancer cell comprises an activated ERBB2,comprising administering to a subject in need thereof an effectiveamount of a compound disclosed herein, or a pharmaceutically acceptablesalt, hydrate, solvate, or stereoisomer thereof. In certain embodiments,the ERBB2 activation is selected from mutation of ERBB2, expression ofERBB2 and amplification of ERBB2. In further embodiments, the mutationis a mutation in exon 20 of ERBB2.

In another aspect, the application provides a method of treating orpreventing cancer, wherein the cancer cell comprises an activated ERBB2,comprising administering to a subject in need thereof an effectiveamount of a compound disclosed herein, or a pharmaceutically acceptablesalt, hydrate, solvate, or stereoisomer thereof, and a second agent thatprevents ERBB2 dimer formation. In certain embodiments, the ERBB2activation is selected from mutation of ERBB2, expression of ERBB2 andamplification of ERBB2. In further embodiments, the mutation is amutation in exon 20 of ERBB2. In some embodiments, the second agent thatprevents ERBB2 dimer formation is an antibody. In further embodiments,the second agent that prevents ERBB2 dimer formation is cetuximab,trastuzumab, or panitumumab. In further embodiments, the second agentthat prevents ERBB2 dimer formation is cetuximab.

In another aspect, the application provides a method of treating cancerin a subject, wherein the subject is identified as being in need ofERBB2 inhibition for the treatment of cancer, comprising administeringto the subject in need thereof an effective amount of a compounddisclosed herein, or a pharmaceutically acceptable salt, hydrate,solvate, or stereoisomer thereof.

In another aspect, the application provides a method of treating cancerin a subject, wherein the subject is identified as being in need ofERBB2 inhibition for the treatment of cancer, comprising administeringto the subject in need thereof an effective amount of a compounddisclosed herein, or a pharmaceutically acceptable salt, hydrate,solvate, or stereoisomer thereof, and optionally a second agent thatprevents ERBB2 dimer formation. In some embodiments, the second agentthat prevents ERBB2 dimer formation is an antibody. In furtherembodiments, the second agent that prevents ERBB2 dimer formation iscetuximab, trastuzumab, or panitumumab. In further embodiments, thesecond agent that prevents ERBB2 dimer formation is cetuximab.

Another aspect of the application provides a method of preventingresistance to a known EGFR inhibitor, including but not limited to,gefitinib, erlotinib, afatinib, lapatinib, neratinib, WZ4002, CL-387785,AZD9291, and CO-1686, in a disease, comprising administering to asubject in need thereof an effective amount of a compound disclosedherein, or a pharmaceutically acceptable salt, hydrate, solvate, orstereoisomer thereof.

Another aspect of the application provides a method of preventingresistance to a known EGFR inhibitor, including but not limited to,gefitinib, erlotinib, afatinib, lapatinib, neratinib, WZ4002, CL-387785,AZD9291, and CO-1686, in a disease, comprising administering to asubject in need thereof an effective amount of a compound disclosedherein, or a pharmaceutically acceptable salt, hydrate, solvate, orstereoisomer thereof, and a second agent that prevents EGFR dimerformation. In some embodiments, the second agent that prevents EGFRdimer formation is an antibody. In further embodiments, the second agentthat prevents EGFR dimer formation is cetuximab, trastuzumab, orpanitumumab. In further embodiments, the second agent that prevents EGFRdimer formation is cetuximab.

In certain embodiments, the application provides a method of treatingany of the disorders described herein, wherein the subject is a human.In certain embodiments, the application provides a method of preventingany of the disorders described herein, wherein the subject is a human.

In another aspect, the application provides a compound disclosed herein,or a pharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof for use in the manufacture of a medicament for treating orpreventing a disease in which EGFR plays a role.

In another aspect, the application provides a compound disclosed herein,or a pharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof, and a second agent that prevents EGFR dimer formation for usein the manufacture of a medicament for treating or preventing a diseasein which EGFR plays a role. In some embodiments, the second agent thatprevents EGFR dimer formation is an antibody. In further embodiments,the second agent that prevents EGFR dimer formation is cetuximab,trastuzumab, or panitumumab. In further embodiments, the second agentthat prevents EGFR dimer formation is cetuximab.

In still another aspect, the application provides the use of a compounddisclosed herein, or a pharmaceutically acceptable salt, hydrate,solvate, or stereoisomer thereof, in the treatment or prevention of adisease in which EGFR plays a role.

In another aspect, the application provides the use of a compounddisclosed herein, or a pharmaceutically acceptable salt, hydrate,solvate, or stereoisomer thereof, and a second agent that prevents EGFRdimer formation in the treatment or prevention of a disease in whichEGFR plays a role. In some embodiments, the second agent that preventsEGFR dimer formation is an antibody. In further embodiments, the secondagent that prevents EGFR dimer formation is cetuximab, trastuzumab, orpanitumumab. In further embodiments, the second agent that prevents EGFRdimer formation is cetuximab.

As inhibitors of EGFR kinases, the compounds and compositions of thisapplication are particularly useful for treating or lessening theseverity of a disease, condition, or disorder where a protein kinase isimplicated in the disease, condition, or disorder. In one aspect, thepresent application provides a method for treating or lessening theseverity of a disease, condition, or disorder where a protein kinase isimplicated in the disease state. In another aspect, the presentapplication provides a method for treating or lessening the severity ofa kinase disease, condition, or disorder where inhibition of enzymaticactivity is implicated in the treatment of the disease. In anotheraspect, this application provides a method for treating or lessening theseverity of a disease, condition, or disorder with compounds thatinhibit enzymatic activity by binding to the protein kinase. Anotheraspect provides a method for treating or lessening the severity of akinase disease, condition, or disorder by inhibiting enzymatic activityof the kinase with a protein kinase inhibitor.

In some embodiments, said method is used to treat or prevent a conditionselected from autoimmune diseases, inflammatory diseases, proliferativeand hyperproliferative diseases, immunologically-mediated diseases, bonediseases, metabolic diseases, neurological and neurodegenerativediseases, cardiovascular diseases, hormone related diseases, allergies,asthma, and Alzheimer's disease. In other embodiments, said condition isselected from a proliferative disorder and a neurodegenerative disorder.

One aspect of this application provides compounds that are useful forthe treatment of diseases, disorders, and conditions characterized byexcessive or abnormal cell proliferation. Such diseases include, but arenot limited to, a proliferative or hyperproliferative disease, and aneurodegenerative disease. Examples of proliferative andhyperproliferative diseases include, without limitation, cancer. Theterm “cancer” includes, but is not limited to, the following cancers:breast ovary; cervix; prostate; testis, genitourinary tract; esophagus;larynx, glioblastoma; neuroblastoma; stomach; skin, keratoacanthoma;lung, epidermoid carcinoma, large cell carcinoma, small cell carcinoma,lung adenocarcinoma; bone; colon; colorectal; adenoma; pancreas,adenocarcinoma; thyroid, follicular carcinoma, undifferentiatedcarcinoma, papillary carcinoma; seminoma; melanoma: sarcoma: bladdercarcinoma; liver carcinoma and biliary passages; kidney carcinoma;myeloid disorders; lymphoid disorders, Hodgkin's, hairy cells; buccalcavity and pharynx (oral), lip, tongue, mouth, pharynx; small intestine;colonrectum, large intestine, rectum, brain and central nervous system;chronic myeloid leukemia (CML), and leukemia. The term “cancer”includes, but is not limited to, the following cancers: myeloma,lymphoma, or a cancer selected from gastric, renal, or and the followingcancers: head and neck, oropharangeal, non-small cell lung cancer(NSCLC), endometrial, hcpatocarcinoma, Non-Hodgkins lymphoma, andpulmonary.

The term “cancer” refers to any cancer caused by the proliferation ofmalignant neoplastic cells, such as tumors, neoplasms, carcinomas,sarcomas, leukemias, lymphomas and the like. For example, cancersinclude, but are not limited to, mesothelioma, leukemias and lymphomassuch as cutaneous T-cell lymphomas (CTCL), noncutaneous peripheralT-cell lymphomas, lymphomas associated with human T-cell lymphotrophicvirus (HTLV) such as adult T-cell leukemia lymphoma (ATLL), B-celllymphoma, acute nonlymphocytic leukemias, chronic lymphocyte leukemia,chronic myelogenous leukemia, acute myelogenous leukemia, lymphomas, andmultiple myeloma, non-Hodgkin lymphoma, acute lymphatic leukemia (ALL),chronic lymphatic leukemia (CLL), Hodgkin's lymphoma, Burkitt lymphoma,adult T-cell leukemia lymphoma, acute-myeloid leukemia (AML), chronicmyeloid leukemia (CML), or hepatocellular carcinoma. Further examplesinclude myelodisplastic syndrome, childhood solid tumors such as braintumors, neuroblastoma, retinoblastoma, Wilms' tumor, bone tumors, andsoft-tissue sarcomas, common solid tumors of adults such as head andneck cancers (e.g., oral, laryngeal, nasopharyngeal and esophageal),genitourinary cancers (e.g., prostate, bladder, renal, uterine, ovarian,testicular), lung cancer (e.g., small-cell and non-small cell), breastcancer, pancreatic cancer, melanoma and other skin cancers, stomachcancer, brain tumors, tumors related to Gorlin's syndrome (e.g.,medulloblastoma, meningioma, etc.), and liver cancer. Additionalexemplary forms of cancer which may be treated by the subject compoundsinclude, but are not limited to, cancer of skeletal or smooth muscle,stomach cancer, cancer of the small intestine, rectum carcinoma, cancerof the salivary gland, endometrial cancer, adrenal cancer, anal cancer,rectal cancer, parathyroid cancer, and pituitary cancer.

Additional cancers that the compounds described herein may be useful inpreventing, treating and studying are, for example, colon carcinoma,familiary adenomatous polyposis carcinoma and hereditary non-polyposiscolorectal cancer, or melanoma. Further, cancers include, but are notlimited to, labial carcinoma, larynx carcinoma, hypopharynx carcinoma,tongue carcinoma, salivary gland carcinoma, gastric carcinoma,adenocarcinoma, thyroid cancer (medullary and papillary thyroidcarcinoma), renal carcinoma, kidney parenchyma carcinoma, cervixcarcinoma, uterine corpus carcinoma, endometrium carcinoma, chorioncarcinoma, testis carcinoma, urinary carcinoma, melanoma, brain tumorssuch as glioblastoma, astrocytoma, meningioma, medulloblastoma andperipheral neuroectodermal tumors, gall bladder carcinoma, bronchialcarcinoma, multiple myeloma, basalioma, teratoma, retinoblastoma,choroidea melanoma, seminoma, rhabdomyosarcoma, craniopharyngeoma,osteosarcoma, chondrosarcoma, myosarcoma, liposarcoma, fibrosarcoma,Ewing sarcoma, and plasmocytoma. In one aspect of the application, thepresent application provides for the use of one or more compounds of theapplication in the manufacture of a medicament for the treatment ofcancer, including without limitation the various types of cancerdisclosed herein.

In some embodiments, the compounds of this application are useful fortreating cancer, such as colorectal, thyroid, breast, and lung cancer:and myeloproliferative disorders, such as polycythemia vera,thrombocythemia, myeloid metaplasia with myelofibrosis, chronicmyelogenous leukemia, chronic myelomonocytic leukemia, hypereosinophilicsyndrome, juvenile myelomonocytic leukemia, and systemic mast celldisease. In some embodiments, the compounds of this application areuseful for treating hematopoietic disorders, in particular,acute-myelogenous leukemia (AML), chronic-myelogenous leukemia (CML),acute-promyelocytic leukemia, and acute lymphocyte leukemia (ALL).

This application further embraces the treatment or prevention of cellproliferative disorders such as hyperplasias, dysplasias andpre-cancerous lesions. Dysplasia is the earliest form of pre-cancerouslesion recognizable in a biopsy by a pathologist. The subject compoundsmay be administered for the purpose of preventing said hyperplasias,dysplasias or pre-cancerous lesions from continuing to expand or frombecoming cancerous. Examples of pre-cancerous lesions may occur in skin,esophageal tissue, breast and cervical intra epithelial tissue.

Examples of neurodegenerative diseases include, without limitation,Adrenoleukodystrophy (ALD). Alexander's disease, Alper's disease.Alzheimer's disease, Amyotrophic lateral sclerosis (Lou Gehrig'sDisease), Ataxia telangiectasia, Batten disease (also known asSpielmeyer-Vogt-Sjogren-Batten disease), Bovine spongiformencephalopathy (BSE), Canavan disease, Cockayne syndrome, Corticobasaldegeneration, Creutzfeldt-Jakob disease, Familial fatal insomnia,Frontotemporal lobar degeneration, Huntington's disease, HIV-associateddementia, Kennedy's disease, Krabbe's disease. Lewy body dementia,Neuroborreliosis, Machado-Joseph disease (Spinocerebellar ataxia type3), Multiple System Atrophy, Multiple sclerosis, Narcolepsy, NiemannPick disease, Parkinson's disease, Pelizaeus-Merzbacher Disease, Pick'sdisease, Primary lateral sclerosis, Prion diseases, ProgressiveSupranuclear Palsy, Refsum's disease, Sandhoff disease, Schilder'sdisease, Subacute combined degeneration of spinal cord secondary toPernicious Anaemia, Spielmeyer-Vogt-Sjogren-Batten disease (also knownas Batten disease), Spinocerebellar ataxia (multiple types with varyingcharacteristics), Spinal muscular atrophy, Steele-Richardson-Olszewskidisease, Tabes dorsalis, and Toxic encephalopathy.

Another aspect of this application provides a method for the treatmentor lessening the severity of a disease selected from a proliferative orhyperproliterative disease, or a neurodegenerative disease, comprisingadministering an effective amount of a compound, or a pharmaceuticallyacceptable composition comprising a compound, to a subject in needthereof. In other embodiments, the method further comprisesadministering a second agent that prevents EGFR dimer formation. In someembodiments, the second agent that prevents EGFR dimer formation is anantibody. In further embodiments, the second agent that prevents EGFRdimer formation is cetuximab, trastuzumab, or panitumumab. In furtherembodiments, the second agent that prevents EGFR dimer formation iscetuximab.

As inhibitors of EGFR kinases, the compounds and compositions of thisapplication are also useful in biological samples. One aspect of theapplication relates to inhibiting protein kinase activity in abiological sample, which method comprises contacting said biologicalsample with a compound of the application or a composition comprisingsaid compound. The term “biological sample”, as used herein, means an invitro or an ex vivo sample, including, without limitation, cell culturesor extracts thereof; biopsied material obtained from a mammal orextracts thereof; and blood, saliva, urine, feces, semen, tears, orother body fluids or extracts thereof. Inhibition of protein kinaseactivity in a biological sample is useful fora variety of purposes thatare known to one of skill in the art. Examples of such purposes include,but are not limited to, blood transfusion, organ-transplantation, andbiological specimen storage.

Another aspect of this application relates to the study of EGFR kinasesin biological and pathological phenomena; the study of intracellularsignal transduction pathways mediated by such protein kinases; and thecomparative evaluation of new protein kinase inhibitors. Examples ofsuch uses include, but are not limited to, biological assays such asenzyme assays and cell-based assays.

The activity of the compounds and compositions of the presentapplication as EGFR kinase inhibitors may be assayed in vitro, in vivo,or in a cell line. In vitro assays include assays that determineinhibition of either the kinase activity or ATPase activity of theactivated kinase. Alternate in vitro assays quantitate the ability ofthe inhibitor to bind to the protein kinase and may be measured eitherby radio labelling the inhibitor prior to binding, isolating theinhibitor/kinase complex and determining the amount of radio labelbound, or by running a competition experiment where new inhibitors areincubated with the kinase bound to known radioligands. Detailedconditions for assaying a compound utilized in this application as aninhibitor of various kinases are set forth in the Examples below.

In accordance with the foregoing, the present application furtherprovides a method for preventing or treating any of the diseases ordisorders described above in a subject in need of such treatment, whichmethod comprises administering to said subject a therapeuticallyeffective amount of a compound of the application, or a pharmaceuticallyacceptable salt, hydrate, solvate, or stereoisomer thereof, andoptionally a second agent that prevents EGFR dimer formation. For any ofthe above uses, the required dosage will vary depending on the mode ofadministration, the particular condition to be treated and the effectdesired.

In other embodiments, the compound and the second agent that preventsEGFR dimer formation are administered simultaneously or sequentially.

PHARMACEUTICAL COMPOSITIONS

In another aspect, the application provides a pharmaceutical compositioncomprising a compound disclosed herein, or a pharmaceutically acceptablesalt, hydrate, solvate, or stereoisomer thereof, together with apharmaceutically acceptable carrier.

In another aspect, the application provides a pharmaceutical compositioncomprising a compound disclosed herein, or a pharmaceutically acceptablesalt, hydrate, solvate, or stereoisomer thereof, and a second agent thatprevents EGFR dimer formation together with a pharmaceuticallyacceptable carrier. In some embodiments, the second agent that preventsEGFR dimer formation is an antibody. In further embodiments, the secondagent that prevents EGFR dimer formation is cetuximab, trastuzumab, orpanitumumab. In further embodiments, the second agent that prevents EGFRdimer formation is cetuximab.

Compounds of the application can be administered as pharmaceuticalcompositions by any conventional route, in particular enterally, e.g.,orally, e.g., in the form of tablets or capsules, or parenterally, e.g.,in the form of injectable solutions or suspensions, topically, e.g., inthe form of lotions, gels, ointments or creams, or in a nasal orsuppository form. Pharmaceutical compositions comprising a compound ofthe present application in free form or in a pharmaceutically acceptablesalt form in association and optionally a second agent that preventsEGFR dimer formation with at least one pharmaceutically acceptablecarrier or diluent can be manufactured in a conventional manner bymixing, granulating or coating methods. For example, oral compositionscan be tablets or gelatin capsules comprising the active ingredienttogether with a) diluents, e.g., lactose, dextrose, sucrose, mannitol,sorbitol, cellulose and/or glycine; b) lubricants, e.g., silica, talcum,stearic acid, its magnesium or calcium salt and/or polyethyleneglycol;for tablets also c) binders, e.g., magnesium aluminum silicate, starchpaste, gelatin, tragacanth, methylcellulose, sodiumcarboxymethylcellulose and or polyvinylpyrrolidone; if desired d)disintegrants, e.g., starches, agar, alginic acid or its sodium salt, oreffervescent mixtures; and/or e) absorbents, colorants, flavors andsweeteners. Injectable compositions can be aqueous isotonic solutions orsuspensions, and suppositories can be prepared from fatty emulsions orsuspensions. The compositions may be sterilized and/or containadjuvants, such as preserving, stabilizing, wetting or emulsifyingagents, solution promoters, salts for regulating the osmotic pressureand/or buffers. In addition, they may also contain other therapeuticallyvaluable substances. Suitable formulations for transdermal applicationsinclude an effective amount of a compound of the present applicationwith a carrier. A carrier can include absorbable pharmacologicallyacceptable solvents to assist passage through the skin of the host. Forexample, transdermal devices are in the form of a bandage comprising abacking member, a reservoir containing the compound optionally withcarriers, optionally a rate controlling barrier to deliver the compoundto the skin of the host at a controlled and predetermined rate over aprolonged period of time, and means to secure the device to the skin.Matrix transdermal formulations may also be used. Suitable formulationsfor topical application, e.g., to the skin and eyes, are preferablyaqueous solutions, ointments, creams or gels well-known in the art. Suchmay contain solubilizers, stabilizers, tonicity enhancing agents,buffers and preservatives.

Compounds and compositions of the application can be administered intherapeutically effective amounts in a combinational therapy with one ormore therapeutic agents (pharmaceutical combinations) or modalities,e.g., a second agent that prevents EGFR dimer formation, non-drugtherapies, etc. For example, synergistic effects can occur with agentsthat prevents EGFR dimer formation, other anti-proliferative,anti-cancer, immunomodulatory or anti-inflammatory substances. Where thecompounds of the application are administered in conjunction with othertherapies, dosages of the co-administered compounds will of course varydepending on the type of co-drug employed, on the specific drugemployed, on the condition being treated and so forth.

Combination therapy includes the administration of the subject compoundsin further combination with one or more other biologically activeingredients (such as, but not limited to, a second agent that preventsEGFR dimer formation, a second and different antineoplastic agent) andnon-drug therapies (such as, but not limited to, surgery or radiationtreatment). For instance, the compounds of the application can be usedin combination with other pharmaceutically active compounds, preferablycompounds that are able to enhance the effect of the compounds of theapplication. The compounds of the application can be administeredsimultaneously (as a single preparation or separate preparation) orsequentially to the other drug therapy or treatment modality. Ingeneral, a combination therapy envisions administration of two or moredrugs during a single cycle or course of therapy.

In one aspect of the application, the compounds may be administered incombination with one or more agents that prevent EGFR dimer formation.In some embodiments, the second agent that prevents EGFR dimer formationis an antibody. In further embodiments, the second agent that preventsEGFR dimer formation is cetuximab, trastuzumab, or panitumumab. Infurther embodiments, the second agent that prevents EGFR dimer formationis cetuximab.

In another aspect of the application, the compounds may be administeredin combination with one or more separate pharmaceutical agents, e.g., achemotherapeutic agent, an immunotherapeutic agent, or an adjunctivetherapeutic agent. In one embodiment, the chemotherapeutic agent reducesor inhibits the binding of ATP with EGFR (e.g., gefitinib, erlotinib,afatinib, lapatinib, nerabinib, CL-387785, AZD9291, CO-1686 or WZ4002).

The pharmaceutical compositions of the present application comprise atherapeutically effective amount of a compound of the presentapplication formulated together with one or more pharmaceuticallyacceptable carriers. As used herein, the term “pharmaceuticallyacceptable carrier” means a non-toxic, inert solid, semi-solid or liquidfiller, diluent, encapsulating material or formulation auxiliary of anytype. The pharmaceutical compositions of this application can beadministered to humans and other animals orally, rectally, parenterally,intracistemally, intravaginally, intraperitoneally, topically (as bypowders, ointments, or drops), buccally, or as an oral or nasal spray.In other embodiments, the composition further comprises administering asecond agent that prevents EGFR dimer formation. In some embodiments,the second agent that prevents EGFR dimer formation is an antibody. Infurther embodiments, the second agent that prevents EGFR dimer formationis cetuximab, trastuzumab, or panitumumab. In further embodiments, thesecond agent that prevents EGFR dimer formation is cetuximab.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, microemulsions, solutions, suspensions, syrups andelixirs. In addition to the active compounds, the liquid dosage formsmay contain inert diluents commonly used in the art such as, forexample, water or other solvents, solubilizing agents and emulsifierssuch as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, dimethylformamide, oils (in particular, cottonseed, groundnut,com, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfurylalcohol, polyethylene glycols and fatty acid esters of sorbitan, andmixtures thereof. Besides inert diluents, the oral compositions can alsoinclude adjuvants such as wetting agents, emulsifying and suspendingagents, sweetening, flavoring, and perfuming agents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolutionwhich, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisapplication with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid compositions of a similar type may also be employed as fillers insoft and hard filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like.

The active compounds can also be in micro-encapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents.

Dosage forms for topical or transdermal administration of a compound ofthis application include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, ear drops, eye ointments, powders and solutionsare also contemplated as being within the scope of this application.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this application, excipients such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to the compounds of thisapplication, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants suchas chlorofluorohydrocarbons.

Transdermal patches have the added advantage of providing controlleddelivery of a compound to the body. Such dosage forms can be made bydissolving or dispensing the compound in the proper medium. Absorptionenhancers can also be used to increase the flux of the compound acrossthe skin. The rate can be controlled by either providing a ratecontrolling membrane or by dispersing the compound in a polymer matrixor gel.

According to the methods of treatment of the present application,disorders are treated or prevented in a subject, such as a human orother animal, by administering to the subject a therapeuticallyeffective amount of a compound of the application, in such amounts andfor such time as is necessary to achieve the desired result. The term“therapeutically effective amount” of a compound of the application, asused herein, means a sufficient amount of the compound so as to decreasethe symptoms of a disorder in a subject. As is well understood in themedical arts a therapeutically effective amount of a compound of thisapplication will be at a reasonable benefit/risk ratio applicable to anymedical treatment.

In general, compounds of the application will be administered intherapeutically effective amounts via any of the usual and acceptablemodes known in the art, either singly or in combination with one or moretherapeutic agents. A therapeutically effective amount may vary widelydepending on the severity of the disease, the age and relative health ofthe subject, the potency of the compound used and other factors. Ingeneral, satisfactory results are indicated to be obtained systemicallyat daily dosages of from about 0.03 to 2.5 mg/kg per body weight. Anindicated daily dosage in the larger mammal, e.g., humans, is in therange from about 0.5 mg to about 100 mg, conveniently administered,e.g., in divided doses up to four times a day or in retard form.Suitable unit dosage forms for oral administration comprise from ca. 1to 50 mg active ingredient.

In certain embodiments, a therapeutic amount or dose of the compounds ofthe present application may range from about 0.1 mg/Kg to about 500mg/Kg, alternatively from about 1 to about 50 mg/Kg. In general,treatment regimens according to the present application compriseadministration to a patient in need of such treatment from about 10 mgto about 1000 mg of the compound(s) of this application per day insingle or multiple doses. Therapeutic amounts or doses will also varydepending on route of administration, as well as the possibility ofco-usage with other agents.

Upon improvement of a subject's condition, a maintenance dose of acompound, composition or combination of this application may beadministered, if necessary. Subsequently, the dosage or frequency ofadministration, or both, may be reduced, as a function of the symptoms,to a level at which the improved condition is retained when the symptomshave been alleviated to the desired level, treatment should cease. Thesubject may, however, require intermittent treatment on a long-termbasis upon any recurrence of disease symptoms.

It will be understood, however, that the total daily usage of thecompounds and compositions of the present application will be decided bythe attending physician within the scope of sound medical judgment. Thespecific inhibitory dose for any particular patient will depend upon avariety of factors including the disorder being treated and the severityof the disorder; the activity of the specific compound employed; thespecific composition employed; the age, body weight, general health, sexand diet of the patient; the time of administration, route ofadministration, and rate of excretion of the specific compound employed;the duration of the treatment; drugs used in combination or coincidentalwith the specific compound employed; and like factors well known in themedical arts.

The application also provides for a pharmaceutical combinations, e.g., akit, comprising a) a first agent which is a compound of the applicationas disclosed herein, in free form or in pharmaceutically acceptable saltform, and b) at least one co-agent. The kit can comprise instructionsfor its administration.

The terms “co-administration” or “combined administration” or the likeas utilized herein are meant to encompass administration of the selectedtherapeutic agents to a single patient, and are intended to includetreatment regimens in which the agents are not necessarily administeredby the same route of administration or at the same time.

The term “pharmaceutical combination” as used herein means a productthat results from the mixing or combining of more than one activeingredient and includes both fixed and non-fixed combinations of theactive ingredients. The term “fixed combination” means that the activeingredients, e.g., a compound of the application and a co-agent, areboth administered to a patient simultaneously in the form of a singleentity or dosage. The term “non-fixed combination” means that the activeingredients, e.g., a compound of the application and a co-agent, areboth administered to a patient as separate entities eithersimultaneously, concurrently or sequentially with no specific timelimits, wherein such administration provides therapeutically effectivelevels of the two compounds in the body of the patient. The latter alsoapplies to cocktail therapy, e.g., the administration of three or moreactive ingredients.

In certain embodiments, these compositions optionally further compriseone or more additional therapeutic agents. For example, an agent thatprevents EGFR dimer formation, chemotherapeutic agents or otherantiproliferative agents may be combined with the compounds of thisapplication to treat proliferative diseases and cancer.

Some examples of materials which can serve as pharmaceuticallyacceptable carriers include, but are not limited to, ion exchangers,alumina, aluminum stearate, lecithin, scrum proteins, such as humanserum albumin, buffer substances such as phosphates, glycine, sorbicacid, or potassium sorbate, partial glyceride mixtures of saturatedvegetable fatty acids, water, salts or electrolytes, such as protaminesulfate, disodium hydrogen phosphate, potassium hydrogen phosphate,sodium chloride, zinc salts, colloidal silica, magnesium trisilicate,polyvinyl pyrrolidone, polyacrylates, waxes,polyethylenepolyoxypropylene-block polymers, wool fat, sugars such aslactose, glucose and sucrose; starches such as corn starch and potatostarch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethyl cellulose and cellulose acetate; powdered tragacanth;malt: gelatin; talc; excipients such as cocoa butter and suppositorywaxes, oils such as peanut oil, cottonseed oil; safflower oil; sesameoil; olive oil; corn oil and soybean oil; glycols; such a propyleneglycol or polyethylene glycol; esters such as ethyl oleate and ethyllaurate, agar; buffering agents such as magnesium hydroxide and aluminumhydroxide; alginic acid; pyrogen-free water, isotonic saline; Ringer'ssolution; ethyl alcohol, and phosphate buffer solutions, as well asother non-toxic compatible lubricants such as sodium lauryl sulfate andmagnesium stearate, as well as coloring agents, releasing agents,coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the composition,according to the judgment of the formulator. The protein kinaseinhibitors or pharmaceutical salts thereof may be formulated intopharmaceutical compositions for administration to animals or humans.These pharmaceutical compositions, which comprise an amount of theprotein inhibitor effective to treat or prevent a proteinkinase-mediated condition and a pharmaceutically acceptable carrier, areother embodiments of the present application.

In another aspect, the application provides a kit comprising a compoundcapable of inhibiting kinase activity selected from one or morecompounds of disclosed herein, or pharmaceutically acceptable salts,hydrates, solvates, prodrugs, stereoisomers, or tautomers thereof, andinstructions for use in treating cancer. In certain embodiments, the kitfurther comprises components for performing a test to determine whethera subject has activating and/or drug resistance mutations in EGFR.

In another aspect, the application provides a kit comprising a compoundcapable of inhibiting EGFR activity selected from a compound disclosedherein, or a pharmaceutically acceptable salt, hydrate, solvate, orstereoisomer thereof.

In another aspect, the application provides a kit comprising a compoundcapable of inhibiting kinase activity selected from one or morecompounds of disclosed herein, or pharmaceutically acceptable salts,hydrates, solvates, prodrugs, stereoisomers, or tautomers thereof, asecond agent that prevents EGFR dimer formation, and instructions foruse in treating cancer. In certain embodiments, the kit furthercomprises components for performing a test to determine whether asubject has activating and/or drug resistance mutations in EGFR. In someembodiments, the second agent that prevents EGFR dimer formation is anantibody. In further embodiments, the second agent that prevents EGFRdimer formation is cetuximab, trastuzumab, or panitumumab. In furtherembodiments, the second agent that prevents EGFR dimer formation iscetuximab.

In another aspect, the application provides a kit comprising a compoundcapable of inhibiting EGFR activity selected from a compound disclosedherein, or a pharmaceutically acceptable salt, hydrate, solvate, orstereoisomer thereof and second agent wherein the second agent preventsEGFR dimer formation. In some embodiments, the second agent thatprevents EGFR dimer formation is an antibody. In further embodiments,the second agent that prevents EGFR dimer formation is cetuximab,trastuzumab, or panitumumab. In further embodiments, the second agentthat prevents EGFR dimer formation is cetuximab.

The application is further illustrated by the following examples andsynthesis schemes, which are not to be construed as limiting thisapplication in scope or spirit to the specific procedures hereindescribed. It is to be understood that the examples are provided toillustrate certain embodiments and that no limitation to the scope ofthe application is intended thereby. It is to be further understood thatresort may be had to various other embodiments, modifications, andequivalents thereof which may suggest themselves to those skilled in theart without departing from the spirit of the present application and/orscope of the appended claims.

EXAMPLES Analytical Methods, Materials, and Instrumentation

Starting materials, reagents and solvents were purchased from commercialsuppliers and were used without further purification unless otherwisenoted. All reactions were monitored using a Waters Acquity UPLC/MSsystem (Waters PDA eλ Detector, QDa Detector, Sample manager—FL, BinarySolvent Manager) using Acquit), UPLC® BEH C18 column (2.1×50 mm, 1.7 μmparticle size): solvent gradient=85% A at 0 min, 1% A at 1.6 min;solvent A=0.1% formic acid in Water; solvent B=0.1% formic acid inAcetonitrile; flow rate: 0.6 mL/min. Reaction products were purified byflash column chromatography using CombiFlash® Rf with Teledyne IscoRediSep® R_(f) columns (4 g, 12 g, 24 g, 40 g, or 80 g) and Waters HPLCsystem using SunFire™ Prep C18 column (19×100 mm, 5 μm particle size):solvent gradient=80% A at 0 min, 10% A at 25 min; solvent A=0.035% TFAin Water; solvent B=0.035% TFA in MeOH; flow rate: 25 mL/min. ¹H NMRspectra were recorded on 500 MHz Bruker Avance III spectrometers.Chemical shifts are reported relative to methanol (δ=3.30), chloroform(δ=7.24) or dimethyl sulfoxide (δ=2.50) for ¹H NMR and ¹³C NMR. Data arereported as (br=broad, s=singlet, d=doublet, t=triplet, q=quartet,m=multiplet).

Abbreviations used in the following examples and elsewhere herein are:

-   -   anh. anhydrous    -   atm atmosphere    -   Bn benzyl    -   br broad    -   brsm based on recovered starting material    -   brine NaCl (sat.)    -   CDI carbonyldiimidazole    -   CHCI₃ chloroform    -   DCM dichloromethane    -   DIEA N,N-diisopropylethylamine    -   DIPEA N,N-diisopropylethylamine    -   DMA N,N-dimethylacetamide    -   DMF N,N-dimethylformamide    -   DMSO dimethyl sulfoxide    -   EDCI N-(3-dimethylaminopropyl)-N′-ethylcarbondiimide        hydrochloride    -   ESI electrospray ionization    -   EtOAc ethyl acetate    -   EtOH ethanol    -   Fe iron (powder)    -   HCl hydrochloric acid    -   h hour(s)    -   HATU        bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium        3-oxide hexafluoro-phosphate    -   Hex hexanes    -   HOBt 1-hydroxybenzotriazole    -   HPLC high-performance liquid chromatography    -   RP-HPLC reversed phase high-performance liquid chromatography    -   ^(i)PrOH isopropanol    -   LCMS liquid chromatography-mass spectrometry    -   LiHMDS lithium hexamethyldisilazide    -   m multiplet    -   MeOH methanol    -   MHz megahertz    -   min minutes    -   mL milliliter    -   mmol millimole    -   MS mass spectrometry    -   NaHCO₃ sodium bicarbonate    -   Na₂CO₃ sodium carbonate    -   Na₂SO₄ sodium sulfate    -   NH₄Cl ammonium chloride    -   N₂ nitrogen    -   NMR nuclear magnetic resonance    -   Pd₂(dba)₃ tris(dibenzylideneacetone)dipalladium(0)    -   PdCl₂(dppf)₂ [1,1′-bis(diphenylphosphino)ferrocene]palladium(II)        dichloride    -   ppm parts per million    -   rt room temperature ˜25° C.    -   sat. saturated, aqueous    -   TEA trimethylamine    -   TFA trifluoroacetic acid    -   THE tetrahydrofuran    -   TLC thin layer chromatography    -   XPhos 2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl

Example 1: Synthesis of Intermediate I-A(S)-3-amino-1-benzyl-8-fluoro-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-onehydrochloride (Intermediate I-A)

Intermediate I-A for the preparation of the compounds of the applicationcan be synthesized according to the procedures below.

Step 1:(S)-2-((tert-butoxycarbonyl)amino)-3-((4-fluoro-2-nitrophenyl)amino)propanoicacid (1)

To a solution of 1,4-difluoro-2-nitrobenzene (3.0 g, 18.9 mmol) and(S)-3-amino-2-((tert-butoxycarbonyl)amino)propanoic acid (4.2 g, 20.8mmol) in DMF (40 mL) was added NaHCO₃(2.4 g, 28.3 mmol). After stirringat 80° C. for 6 h, the mixture was diluted with EtOAc and filteredthrough celite. The filtrate was washed with 0.5 N HCl, dried overNa₂SO₄, and filtered. The filtrate was concentrated under reducedpressure to obtain(S)-2-((tert-butoxycarbonyl)amino)-3-((4-fluoro-2-nitrophenyl)amino)propanoicacid (1) which was used next step without further purification.

Step 2:(S)-3-((2-amino-4-fluorophenyl)amino)-2-((tert-butoxycarbonyl)amino)propanoicacid (2)

To a solution of the crude(S)-2-((tert-butoxycarbonyl)amino)-3-((4-fluoro-2-nitrophenyl)amino)propanoicacid (1) in THF:EtOH (20 mL:5 mL) were added iron (Fe) powder (1 g) andsat. ammonium chloride (5 mL). After stirring at 50° C. for 2 h, thereaction mixture was filtered through celite and concentrated. Theresidue was re-dissolve in CHCl₃:^(i)PrOH (4:1) and washed with brine.The aqueous layer was further washed with CHCl₃:^(i)PrOH (4:1) twice andthe combined organic layer was dried over Na₂SO₄ and filtered. Thefiltrate was concentrated under reduced pressure to obtain(5)-3-((2-amino-4-fluorophenyl)amino)-2-((tert-butoxycarbonyl)amino)propanoicacid (2) which was used next step without further purification.

Step 3: (S)-tert-butyl(8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)carbamate(3)

To a solution of(S)-3-((2-amino-4-fluorophenyl)amino)-2-((tert-butoxycarbonyl)amino)propanoicacid (2) (1.5 g, 4.79 mmol) in DMF (160 mL) were added HATU (2.7 g, 7.18mmol) and DIPEA (3.3 mL, 19.2 mmol). After stirring for 4 h, the mixturewas diluted with EtOAc and washed with brine (×1) and water (×5). Theorganic layer was dried over Na₂SO₄, filtered, and concentrated underreduced pressure. The residue was purified by flash columnchromatography (Hex:EtOAc=80:20 to 20:80) to give tert-butyl(S)-(8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)carbamate(3) (1.5 g, 27%, three steps) as a dark brown solid.

Step 4: (S)-tert-butyl(1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)carbamate(4)

To a solution of tert-butyl(S)-(8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)carbamate(1.52 g, 5.15 mmol) in anhydrous THE (20 mL) was added LiHMDS (5.67 mL,1.0 M in THF) at −78° C. under nitrogen atmosphere. The resultingsolution was stirred at −78° C. for 30 min, after which benzyl bromide(613 μL, 5.15 mmol) was added dropwise. The resulting reaction mixturewas stirred for 16 h, allowing the temperature to gradually rise to roomtemperature. The reaction mixture was diluted with an excess of EtOAcand washed with sat. NaHCO₃. The organic layer was dried over Na₂SO₄,filtered, and concentrated. The residue was purified by flash columnchromatography (EtOAc:Hex=0:100 to 100:0) to give tert-butyl(S)-(1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)carbamate(4) (1.10 g, 55%, 70% brsm) as a light yellow oil.

Step 5:(S)-3-amino-1-benzyl-8-fluoro-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-onehydrochloride (Intermediate I-A)

To a solution of tert-butyl(S)-(1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)carbamate(4) (1.10 g, 2.85 mmol) in 1,4-dioxane (5.5 mL) was added hydrochloricacid (8.5 mL, 4.0 M in 1,4-dioxane). The resulting reaction mixture wasstirred for 1.5 h, after which the solution was concentrated. Theresidue was co-evaporated four times with DCM:Hex to afford(S)-3-amino-1-benzyl-8-fluoro-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-onehydrochloride (Intermediate I-A) (860 mg, 94%) as an off-white solid.

Example 2: Synthesis of Compound I-1

(S)-3-Amino-1-benzyl-8-fluoro-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-onehydrochloride (Intermediate I-A) (48 mg, 0.15 mmol),4-(4-methylpiperazin-1-yl)benzoic acid (5) (41 mg, 0.19 mmol),N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (43 mg,0.22 mmol), 1-hydroxybenzotriazole (20 mg, 0.15 mmol), andN,N-diisopropylethylamine (105 μL, 0.60 mmol) were dissolved inanhydrous DMF (1.0 mL) and stirred at room temperature for 4 h. Themixture was purified by preparative RP-HPLC to afford(S)—N-(1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)-4-(4-methylpiperazin-1-yl)benzamide(Compound I-1) (49 mg, 67%) as a white solid.

¹H NMR (500 MHz, DMSO-d6) δ 8.38 (d, J=8.2 Hz, 1H), 7.79 (d, J=9.2 Hz,2H), 7.31-7.21 (m, 5H), 7.19-7.14 (m, 1H), 7.04 (dd, J=6.0, 8.7 Hz, 1H),6.99 (d, J=9.2 Hz, 2H), 6.93 (td, J=3.1, 8.4 Hz, 1H), 5.29 (d, J=16.5Hz, 1H), 5.19 (d, J=4.3 Hz, 1H), 4.94 (d, J=16.2 Hz, 1H), 4.79 (dt,J=7.6, 12.1 Hz, 1H), 3.71 (dd, J=9.3, 12.1 Hz, 1H), 3.59 (ddd, J=4.6,6.9, 9.5 Hz, 1H), 3.47-3.34 (m, 4H), 2.91-2.67 (m, 4H), 2.44 (br s, 3H);LCMS (ES) m/z 487.94 [M+H]⁺.

Example 3: Synthesis of Compound I-2

To a solution of(S)-3-amino-1-benzyl-8-fluoro-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-onehydrochloride (Intermediate I-A) (64 mg, 0.20 mmol) in anhydrous DMSO (2mL) was added triethylamine (84 μL, 0.60 mmol) andN,N′-carbonyldiimidazole (32 mg, 0.20 mmol). The resulting solution wasstirred for 15 min, after which 4-(4-methylpiperazin-1-yl)aniline (6)(115 mg, 0.60 mmol) was added. The resulting reaction mixture wasstirred for 16 h and then purified by preparative RP-HPLC to afford(S)-1-(1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)-3-(4-(4-methylpiperazin-1-yl)phenyl)urea(Compound I-2) (37 mg, 37%) as a white solid.

¹H NMR (500 MHz, DMSO-d6) δ 8.62 (s, 1H), 7.27-7.15 (m, 8H), 7.03 (dd,J=6.1, 8.9 Hz, 1H), 6.92 (td, J=2.7, 8.4 Hz, 1H), 6.84 (d, J=9.2 Hz,2H), 6.47 (d, J=7.9 Hz, 1H), 5.34 (d, J=16.2 Hz, 1H), 5.23 (d, J=4.9 Hz,1H), 4.92 (d, J=16.2 Hz, 1H), 4.58-4.51 (m, 1H), 3.71-3.64 (m, 1H),3.34-3.29 (m, 1H), 3.23-3.02 (m, 4H), 2.93-2.69 (m, 4H), 2.47 (br s,3H); LCMS (EST) m/z 502.91 [M+H]⁺.

Example 4: Synthesis of Compound I-3

(S)—N-(1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)-2-(4-morpholinophenyl)acetamide(Compound I-3) was synthesized from(S)-3-amino-1-benzyl-8-fluoro-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-onehydrochloride (Intermediate I-A) and 2-(4-morpholinophenyl)acetic acid(7) by following an analogous procedure to the one outlined for(S)—N-(1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)-4-(4-methylpiperazin-1-yl)benzamide(Compound I-1) in Example 2 above (57 mg, 60%) as off-white solid.

¹H NMR (500 MHz, DMSO-d₆) δ 8.36 (d, J=8.2 Hz, 1H), 7.25-7.20 (m, 5H),7.19-7.14 (m, 1H), 7.11 (d, J=8.5 Hz, 2H), 7.00 (dd, J=6.0, 8.7 Hz, 1H),6.92-6.88 (m, 1H), 6.87 (d, J=8.5 Hz, 2H), 5.31 (d, J=16.2 Hz, 1H), 5.16(br s, 1H), 4.88 (d, J=15.9 Hz, 1H), 4.60-4.53 (m, 1H), 3.74-3.71 (m,4H), 3.57-3.52 (m, 1H), 3.44-3.38 (m, 1H), 3.37 (s, 2H), 3.07-3.05 (m,4H); LCMS (ESI) m/z 488.92 [M+H]⁺.

Example 5: Synthesis of Compound I-4

(S)—N-(1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)-3-(4-methylpiperazin-1-yl)benzamide(Compound I-4) was synthesized from(S)-3-amino-1-benzyl-8-fluoro-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-onehydrochloride (Intermediate I-A) and 3-(4-methylpiperazin-1-yl)benzoicacid (8) by following an analogous procedure to the one outlined for(S)N-(1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)-4-(4-methylpiperazin-1-yl)benzamide(Compound I-1) in Example 2 above (52 mg, 55%) as white solid.

¹H NMR (500 MHz, DMSO-d6) δ 8.62 (d, J=7.9 Hz, 1H), 7.44-7.41 (m, 1H),7.36-7.22 (m, 7H), 7.19-7.15 (m, 2H), 7.05 (dd, J=6.0, 8.7 Hz, 1H), 6.94(td, J=2.9, 8.5 Hz, 1H), 5.30 (d, J=16.2 Hz, 1H), 5.22 (d, J=4.3 Hz,1H), 4.94 (d, J=16.2 Hz, 1H), 4.80 (dt, J=7.6, 11.9 Hz, 1H), 3.72 (dd,J=9.6, 11.7 Hz, 1H), 3.62 (ddd, J=4.4, 7.2, 9.5 Hz, 1H), 3.50-3.32 (m,4H), 3.17-2.95 (m, 4H), 2.67 (br s, 3H); LCMS (ESI) m/z 487.90 [M+H]⁺.

Example 6: Synthesis of Compound I-5

(S)—N-(1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)-2-methoxy-4-morpholinobenzamide(Compound I-5) was synthesized from(S)-3-amino-1-benzyl-8-fluoro-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-onehydrochloride (Intermediate I-A) and 2-methoxy-4-morpholinobenzoic acid(9) by following an analogous procedure to the one outlined for(S)—N-(1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)-4-(4-methylpiperazin-1-yl)benzamide(Compound I-1) in Example 2 above (27 mg, 27%) as white solid.

¹H NMR (500 MHz, DMSO-d₆) δ 8.61 (d, J=7.0 Hz, 1H), 7.75 (d, J=8.9 Hz,1H), 7.29-7.23 (m, 5H), 7.19-7.15 (m, 1H), 7.04 (dd, J=5.8, 8.9 Hz, 1H),6.94 (td, J=2.9, 8.5 Hz, 1H), 6.61-6.56 (m, 2H), 5.39 (d, J=16.2 Hz,1H), 5.30 (d, J=4.9 Hz, 1H), 4.91 (d, J=16.2 Hz, 1H), 4.81 (dt, J=6.7,11.3 Hz, 1H), 3.96 (s, 3H), 3.81 (dt, J=5.6, 9.8 Hz, 1H), 3.75-3.71 (m,4H), 3.42-3.36 (m, 1H), 3.28-3.24 (m, 4H); LCMS (ESI) m/z 504.88 [M+H]⁺.

Example 7: Synthesis of Compound I-6

(S)—N-(1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)-4-(4-methylpiperazin-1-yl)picolinamide(Compound I-6) was synthesized from(S)-3-amino-1-benzyl-8-fluoro-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-onehydrochloride (Intermediate I-A) and 4-(4-methylpiperazin-1-yl)picolinicacid (10) by following an analogous procedure to the one outlined for(S)—N-(1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)-4-(4-methylpiperazin-1-yl)benzamide(Compound I-1) in Example 2 above (15 mg, 15%) as white solid.

¹H NMR (500 MHz, DMSO-d6) δ 8.84 (d, J=7.6 Hz, 1H), 8.28 (d, J=5.8 Hz,1H), 7.44 (d, J=2.7 Hz, 1H), 7.29-7.23 (m, 5H), 7.20-7.16 (m, 1H),7.07-7.04 (m, 2H), 6.95 (td, J=2.9, 8.5 Hz, 1H), 5.34 (d, J=16.2 Hz,1H), 5.30 (d, J=4.9 Hz, 1H), 4.95 (d, J=16.2 Hz, 1H), 4.76 (dt, J=7.2,11.6 Hz, 1H), 3.79-3.74 (m, 1H), 3.60-3.40 (m, 5H), 2.94-2.74 (m, 4H),2.51 (br s, 3H); LCMS (ESI) m/z 488.92 [M+H]⁺.

Example 8: Synthesis of Compounds I-7 and I-15 Step 1:(S)—N-(1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)-5-bromo-2-methoxybenzamide(I-15)

(S)-3-Amino-1-benzyl-8-fluoro-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-onehydrochloride (Intermediate I-A) (128 mg, 0.40 mmol),5-bromo-2-methoxybenzoic acid (11) (116 mg, 0.50 mmol),N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (115 mg,0.60 mmol), 1-hydroxybenzotriazole (54 mg, 0.40 mmol), andN,N-diisopropylethylamine (279 μL, 1.60 mmol) were dissolved inanhydrous DMF (2.0 mL) and stirred at room temperature for 15 min. Themixture was precipitated by addition of an excess of water. Theresulting precipitate was filtered over celite, washed repeatedly withwater, re-dissolved in DCM, and concentrated. The residue was purifiedby flash column chromatography (EtOAc:Hex=0:100 to 100:0) to give(S)—N-(1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)-5-bromo-2-methoxybenzamide(Compound I-15) (147 mg, 74%) as a white solid.

Step 2: tert-butyl(S)-4-(3-((1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)carbamoyl)-4-methoxyphenyl)-3,6-dihydropyridine-1(2H)-carboxylate(14)

A mixture of(S)—N-(1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)-5-bromo-2-methoxybenzamide(Compound I-15) (147 mg, 0.29 mmol), tert-butyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate(13) (269 mg, 0.87 mmol) and a 2 N aqueous solution of sodium carbonate(725 μL, 1.45 mmol) in 1,4-dioxane (4 mL) was degassed by nitrogenbubbling for 10 min and heated to 100° C. Then, PdCl₂(dppf)₂ (24 mg, 29μmol) and XPhos (21 mg, 44 μmol) were added and the resulting reactionmixture was stirred at 100° C. for 30 min. The reaction mixture wascooled to room temperature and filtered through a pad of celite. Thefiltrate was concentrated under reduced pressure and the residue wasre-dissolved in DCM and washed with sat. NaHCO₃. The organic layer wasdried over Na₂SO₄, filtered and concentrated. The resulting residue waspurified by flash column chromatography (EtOAc:Hex=0:100 to 100:0)yielding a semipure tert-butyl(S)-4-(3-((1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)carbamoyl)-4-methoxyphenyl)-3,6-dihydropyridine-1(2H)-carboxylate(14) as an orange oil that was carried forward to the next step.

Step 3:(S)—N-(1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)-2-methoxy-5-(1,2,3,6-tetrahydropyridin-4-yl)benzamide(I-7)

To a solution of tert-butyl(S)-4-(3-((1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)carbamoyl)-4-methoxyphenyl)-3,6-dihydropyridine-1(2H)-carboxylate(14) in dichloromethane (5 mL) was added trifluoroacetic acid (2 mL).The resulting reaction mixture was stirred for 15 min, after which thesolution was concentrated and trifluoroacetic acid was removed underreduced pressure. The residue was purified by preparative RP-HPLC toafford(S)—N-(1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)-2-methoxy-5-(1,2,3,6-tetrahydropyridin-4-yl)benzamide(Compound I-7) (81 mg, 55% over two steps) as a white solid.

¹H NMR (500 MHz, DMSO-d₆) δ 8.85 (br s, 1H), 8.79 (d, J=7.3 Hz, 1H),7.91 (d, J=2.4 Hz, 1H), 7.65 (dd, J=2.6, 8.7 Hz, 1H), 7.30 (dd, J=2.9,10.2 Hz, 1H), 7.28-7.21 (m, 5H), 7.19-7.16 (m, 1H), 7.05 (dd, J=5.8, 8.9Hz, 1H), 6.95 (td, J=2.7, 8.4 Hz, 1H), 6.14-6.11 (m, 1H), 5.39 (d,J=16.2 Hz, 1H), 5.30 (br s, 1H), 4.93 (d, J=16.2 Hz, 1H), 4.82 (dt,J=6.9, 11.4 Hz, 1H), 3.97 (s, 3H), 3.80 (dd, J=6.9, 9.3 Hz, 1H),3.76-3.73 (m, 2H), 3.51-3.44 (m, 1H), 3.35-3.30 (m, J=4.6 Hz, 2H),2.67-2.62 (m, 2H); LCMS (ESI) m/z 500.90 [M+H]⁺.

Example 9: Synthesis of Compound I-8

(S)—N-(1-Benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)-5-morpholinopicolinamide(Compound I-8) was synthesized from(S)-3-amino-1-benzyl-8-fluoro-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-onehydrochloride (Intermediate I-A) and 5-morpholinopicolinic acid (15) byfollowing an analogous procedure to the one outlined for(S)—N-(1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)-4-(4-methylpiperazin-1-yl)benzamide(Compound I-1) in Example 2 above (43 mg, 45%) as white solid.

¹H NMR (500 MHz, DMSO-d₆) δ 8.58 (d, J=7.6 Hz, 1H), 8.35 (d, J=2.7 Hz,1H), 7.82 (d, J=8.5 Hz, 1H), 7.40 (dd, J=3.1, 8.9 Hz, 1H), 7.29-7.22 (m,5H), 7.20-7.15 (m, 1H), 7.05 (dd, J=5.8, 8.9 Hz, 1H), 6.94 (td, J=2.7,8.4 Hz, 1H), 5.35 (d. J=16.2 Hz, 1H), 5.29 (d, J=4.9 Hz, 1H), 4.94 (d,J=16.2 Hz, 1H), 4.80-4.73 (m, 1H), 3.78-3.73 (m, 5H), 3.54 (dd, J=9.6,11.4 Hz, 1H), 3.32-3.29 (m, 4H): LCMS (ESI) m/z 475.92 [M+H]⁺.

Example 10: Synthesis of Compound I-9

(S)-2-(4-(1H-tetrazol-1-yl)phenyl)N-(1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)acetamide(Compound I-9) was synthesized from(S)-3-amino-1-benzyl-8-fluoro-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-onehydrochloride (Intermediate I-A) and2-(4-(1H-tetrazol-1-yl)phenyl)acetic acid (16) by following an analogousprocedure to the one outlined for(S)—N-(1-benzyl-8-fluoro-2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)-4-(4-methylpiperazin-1-y1)benzamide(Compound I-1) in Example 2 above (31 mg, 34%) as white solid.

¹H NMR (500 MHz, DMSO-d₆) δ 10.05 (s, 1H), 8.60 (d, J=7.9 Hz, 1H), 7.82(d, J=8.5 Hz, 2H), 7.52 (d, J=8.5 Hz, 2H), 7.26-7.21 (m, 5H), 7.19-7.14(m, 1H), 7.01 (dd, J=6.1, 8.9 Hz, 1H), 6.91 (td, J=2.7, 8.4 Hz, 1H),5.32 (d, J=16.2 Hz, 1H), 5.18 (br s, 1H), 4.90 (d, J=16.2 Hz, 1H),4.63-4.57 (m, 1H), 3.62-3.56 (m, 3H), 3.44 (dd, J=9.5, 11.9 Hz, 1H):LCMS (ESI) m/z 471.90 [M+H]⁺.

Example 11: Synthesis of Compound I-10(S)-1-benzyl-8-fluoro-3-(6-fluoro-1-oxoisoindolin-2-yl)-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(I-10)

To a solution of(S)-3-amino-1-benzyl-8-fluoro-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-onehydrochloride (Intermediate I-A) (42 mg, 0.11 mmol) and methyl5-fluoro-2-(bromomethyl)benzoate (17) (25 mg, 0.11 mmol) in DMF (1 mL)was added DIEA (115 μl, 0.66 mmol). After stirring at 80° C. for 12 h,the reaction mixture was diluted with EtOAc, washed with bring andwater, dried over sodium sulfate, and filtered. The filtrate wasconcentrated under reduced pressure and purified by prepHPLC to obtain(S)-1-benzyl-8-fluoro-3-(6-fluoro-1-oxoisoindolin-2-yl)-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(Compound I-10) (28 mg, 64%) as a white solid.

¹H NMR (500 MHz, DMSO-d₆) δ 7.75 (dd, J=8.4, 5.0 Hz, 1H), 7.51 (dd,J=8.7, 2.0 Hz, 1H), 7.37-7.29 (m, 2H), 7.27-7.22 (m, 4H), 7.21-7.15 (m,1H), 7.09 (dd, J=8.9, 5.8 Hz, 1H), 6.97 (td, J=8.4, 2.7 Hz, 1H), 5.40(d, J=4.3 Hz, 1H), 5.28 (d, J=16.2 Hz, 1H), 5.09-5.01 (m, 2H), 4.94 (d,J=16.2 Hz, 1H), 4.60 (d, J=17.7 Hz, 1H), 4.01 (d, J=12.5, 9.5 Hz, 1H),3.75-3.67 (m, 1H); LCMS (ESI) m/z 419.88 [M+H]⁺.

Example 12: Synthesis of Compound I-16(S)-1-benzyl-8-fluoro-3-(6-iodo-1-oxoisoindolin-2-yl)-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(I-16)

(S)-1-benzyl-8-fluoro-3-(6-iodo-1-oxoisoindolin-2-yl)-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(Compound I-16) was synthesized from(S)-3-amino-1-benzyl-8-fluoro-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-onehydrochloride (Intermediate I-A) and methyl2-(bromomethyl)-5-iodobenzoate (18) by following an analogous procedureto the one outlined for(S)-1-benzyl-8-fluoro-3-(6-fluoro-1-oxoisoindolin-2-yl)-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(Compound I-10) in Example 11 above.

Example 13: Synthesis of Compound I-11

To a solution of(S)-1-benzyl-8-fluoro-3-(6-iodo-1-oxoisoindolin-2-yl)-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(Compound I-16) (70 mg, 0.132 mmol) and(4-(4-methylpiperazin-1-yl)phenyl)boronic acid (19) (44 mg, 0.198 mmol)in dioxane (1.2 ml) was added 2 N solution of sodium carbonate (0.4 ml,0.792 mmol). The mixture was degassed by sonication for 30 sec andpreheated at 100° C. for 5 min. Then, PdCl₂(dppf)₂ and XPhos were addedto the reaction mixture and stirred at 100° C. for 6 h. After completed,the reaction mixture was cooled to room temperature, diluted with EtOAcand washed with water and brine. The organic layer was dried over sodiumsulfate, filtered and concentrated under reduced pressure. The residuewas purified by prepHPLC to obtain(S)-1-benzyl-8-fluoro-3-(6-(4-(4-methylpiperazin-1-yl)phenyl)-1-oxoisoindolin-2-yl)-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(Compound I-11) (34 mg, 45%) as a white solid.

¹H NMR (500 MHz, DMSO-d₆) δ 7.88 (dd, J=7.9, 1.8 Hz, 1H), 7.85-7.82 (m,1H), 7.66 (d, J=8.2 Hz, 2H), 7.63-7.57 (m, 2H), 7.32 (dd, J=10.4, 2.7Hz, 1H), 7.25 (d, J=4.3 Hz, 4H), 7.21-7.16 (m, 1H), 7.10 (dd, J=8.9, 5.8Hz, 1H), 7.05-7.01 (m, 2H), 6.97 (td, J=8.4, 2.7 Hz, 1H), 5.41 (d, J=4.3Hz, 1H), 5.28 (d, J=16.2 Hz, 1H), 5.10 (d, J=12.5, 6.7 Hz, 1H), 5.06 (d,J=17.4 Hz, 1H), 4.95 (d, J=16.2 Hz, 1H), 4.61 (d, J=17.4 Hz, 1H), 4.05(dd, J=12.5, 9.5 Hz, 1H), 3.77-3.68 (m, 1H), 3.24-3.16 (m, 4H),2.49-2.43 (m, 4H), 2.23 (s, 3H); LCMS (ES) m/z 576.06 [M+H]⁺.

Example 14: Synthesis of Compound I-18

(S)-1-benzyl-8-fluoro-3-(6-(1-methyl-1H-pyrazol-4-yl)-1-oxoisoindolin-2-yl)-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(Compound I-18) was synthesized from(S)-1-benzyl-8-fluoro-3-(6-iodo-1-oxoisoindolin-2-yl)-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(Compound I-16) and (1H-pyrazol-4-yl)boronic acid (20) by following ananalogous procedure to the one outlined for(S)-1-benzyl-8-fluoro-3-(6-(4-(4-methylpiperazin-1-yl)phenyl)-1-oxoisoindolin-2-yl)-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(Compound I-11) in Example 13 above.

¹H NMR (500 MHz, DMSO-d₆) δ 12.98 (br s, 1H), 8.35 (s, 1H), 8.03 (s,1H), 7.92 (s, 1H), 7.90 (dd, J=7.9, 1.8 Hz, 1H), 7.60 (d, J=7.9 Hz, 1H),7.31 (dd, J=10.4, 2.7 Hz, 1H), 7.27-7.22 (m, 4H), 7.21-7.15 (m, 1H),7.10 (dd, J=8.9, 6.1 Hz, 1H), 6.97 (td, J=8.4, 2.7 Hz, 1H), 5.41 (d,J=4.3 Hz, 1H), 5.27 (d, J=16.2 Hz, 1H), 5.10 (d, J=12.4, 6.9 Hz, 1H),5.03 (d, J=17.4 Hz, 1H), 4.96 (d, J=16.2 Hz, 1H), 4.58 (d, J=17.1 Hz,1H), 4.04 (dd, J=12.5, 9.5 Hz, 1H), 3.76-3.68 (m, 1H); LCMS (ESI) m/z467.95 [M+H]⁺.

Example 15: Synthesis of Compound I-13(S)-1-benzyl-8-fluoro-3-(6-(4-methylpiperazin-1-yl)-1-oxoisoindolin-2-yl)-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(I-13)

To a solution of(S)-1-benzyl-8-fluoro-3-(6-iodo-1-oxoisoindolin-2-yl)-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(Compound I-16) (60 mg, 0.114 mmol), 1-methylpiperazine (21) (22.8 mg,0.228 mmol) and potassium carbonate (32 mg, 0.228 mmol) in DMSO wereadded copper(I) iodide (4 mg, 0.023 mmol) and L-proline (4 mg, 0.034mmol). After stirring for 12 hr, the reaction mixture was diluted withEtOAc and washed with sat. NaHCO₃ and brine. The organic layer was driedover sodium sulfate, filtered and concentrated under reduced pressure.The residue was purified by prepHPLC to give(S)-1-benzyl-8-fluoro-3-(6-(4-methylpiperazin-1-yl)-1-oxoisoindolin-2-yl)-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(Compound I-13) (27 mg, 48%) as an off-white solid.

¹H NMR (500 MHz, DMSO-d₆) δ 7.44 (d, J=8.2 Hz, 1H), 7.32-7.22 (m, 6H),7.20-7.14 (m, 1H), 7.13-7.07 (m, 2H), 6.96 (td, J=8.2, 2.8 Hz, 1H), 5.38(d, J=4.3 Hz, 1H), 5.26 (d, J=16.2 Hz, 1H), 5.06 (dd, J=12.5, 6.7 Hz,1H), 4.93 (dd, J=16.3, 14.5 Hz, 2H), 4.45 (d, J=16.8 Hz, 1H), 4.01 (dd,J=12.4, 9.3 Hz, 1H), 3.72-3.66 (m, 1H), 3.20-3.14 (m, 4H), 2.48-2.43 (m,4H), 2.22 (s, 3H); LCMS (ESI) m/z 499.97 [M+H]⁺.

Example 16: Synthesis of Compound I-17(S)-1-benzyl-3-(5-bromo-1-oxoisoindolin-2-yl)-8-fluoro-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one (I-17)

(S)-1-benzyl-3-(5-bromo-1-oxoisoindolin-2-yl)-8-fluoro-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(Compound I-17) was synthesized from(S)-3-amino-1-benzyl-8-fluoro-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-onehydrochloride (Intermediate I A) and methyl4-bromo-2-(bromomethyl)benzoate (22) by following an analogous procedureto the one outlined for(S)-1-benzyl-8-fluoro-3-(6-fluoro-1-oxoisoindolin-2-yl)-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(Compound I-10) in Example 11 above.

Example 17: Synthesis of Compound I-14(S)-1-benzyl-8-fluoro-3-(5-(4-(4-methylpiperazin-1-yl)phenyl)-1-oxoisoindolin-2-yl)-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(I-14)

(S)-1-benzyl-8-fluoro-3-(5-(4-(4-methylpiperazin-1-yl)phenyl)-1-oxoisoindolin-2-yl)-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(Compound I-14) was synthesized from(S)-1-benzyl-3-(5-bromo-1-oxoisoindolin-2-yl)-8-fluoro-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(Compound I-17) and (4-(4-methylpiperazin-1-yl)phenyl)boronic acid (19)by following an analogous procedure to the one outlined for(S)-1-benzyl-8-fluoro-3-(6-(4-(4-methylpiperazin-1-y1)phenyl)-1-oxoisoindolin-2-yl)-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(Compound I-11) in Example 13 above.

¹H NMR (500 MHz, DMSO-d₆) δ 7.84 (s, 1H), 7.75-7.68 (m, 2H), 7.62 (d,J=8.9 Hz, 2H), 7.33 (dd, J=10.2, 2.9 Hz, 1H), 7.25 (d, J=4.6 Hz, 4H),7.21-7.16 (m, 1H), 7.10 (dd, J=8.9, 6.1 Hz, 1H), 7.07-7.03 (m, 2H), 6.97(td, J=8.5, 2.9 Hz, 1H), 5.40 (d, J=4.0 Hz, 1H), 5.30 (d, J=16.2 Hz,1H), 5.12-5.05 (m, 2H), 4.94 (d, J=16.3 Hz, 1H), 4.62 (d, J=17.4 Hz,1H), 4.05 (dd, J=12.5, 9.5 Hz, 1H), 3.76-3.68 (m, 1H), 3.25-3.17 (m,4H), 2.48-2.43 (m, 4H), 2.23 (s, 3H); LCMS (ESI) m/z 576.06 [M+H]⁺.

Example 18: Biochemical/Biological Studies Ba/F3 Cell ProliferationModels

The EGFR mutant L858R, Del E746_A750, L858R/T790M, DelE746_A750/T790M,L858R/T790M/C797S and Del/T790M/C797S Ba/F3 cells were previouslydescribed (Thou et al., Nature 462, (2009), 1070-1074). All cell lineswere maintained in RPMI 1640 (Cellgro; Mediatech Inc., Herndon, Calif.)supplemented with 10% FBS 100 units/mL penicillin, 100 units/mLstreptomycin, and 2 mM glutamine. L858R cells were maintained in ACL-4media (Invitrogen, Carlsbad, Calif.) supplemented with 5% FBS, 100units/mL penicillin, 100 units/mL streptomycin, and 2 mM glutamine. TheEGFR 1941R mutation was introduced via site directed mutagenesis usingthe Quick Change Site-Directed Mutagenesis kit (Stratagene; La Jolla,Calif.) according to the manufacturer's instructions. All constructswere confirmed by DNA sequencing. The constructs were shuttled into theretroviral vector JP1540 using the BD Creator™ System (BD Biosciences).Ba/F3 cells were infected with retrovirus and according to standardprotocols, as described previously (Thou 2009). Stable clones wereobtained by selection in puromycin (2 μg/ml).

Growth and inhibition of growth was assessed by MTS assay and wasperformed according to previously established methods (Zhou 2009). TheMTS assay is a colorimetric method for determining the number of viablecells that is based on the bioreduction of MTS by cells to a formazanproduct that is soluble in cell culture medium and can be detectedspectrophotometrically. Ba/F3 cells of different EGFR genotypes wereexposed to treatment and the number of cells used per experimentdetermined empirically and has been previously established (Zhou 2009).All experimental points were set up in six wells and all experimentswere repeated at least three times. The data was graphically displayedusing GraphPad Prism version 5.0 for Windows (GraphPad Software). Thecurves were fitted using a non-linear regression model with a sigmoidaldose response.

EGFR Protein Expression and Purification

Constructs spanning residues 696-1022 of the human EGFR (including wildtype and L858R, L858R/T790M, T790M, and T790M/V948R mutant sequences)were prepared in a GST-fusion format using the pTriEX system (Novagen)for expression in Sf9 insect cells essentially as described (Yun et al.,PNAS 105, 2070-2075 (2008); Yun et al., Cancer Cell 11, 217-227 (2007)).EGFR kinase proteins were purified by glutathione-affinitychromatography followed by size-exclusion chromatography after cleavagewith TEV or thrombin to remove the GST fusion partner followingestablished procedures. (Yun 2008; Yun 2007).

High-Throughput Screening

Purified EGFR-L858R/T790M enzyme was screened against compounds of thepresent application using HTRF-based biochemical assay format. Thescreening was performed at 1 μM ATP using a single compoundconcentration (12.5 μM), 1322 top hits were picked for follow-up IC₅₀confirmation. IC₅₀ values were determined at both 1 μM and 1 mM ATP toidentify both ATP competitive and non-competitive compounds. Hits werealso counter-screened against wild type EGFR to evaluate the mutantselectivity.

The HTRF-based screen was carried out using 1 μM ATP, and activecompounds were counter-screened at 1 mM ATP and against wild type EGFRto identify those that were potentially non-ATP-competitive and mutantselective. This strategy identified several compounds of distinctchemical classes that were both selective for the L858R/T790M mutantover WT EGFR and relatively insensitive to ATP concentrations,suggesting an allosteric mechanism of action.

HTRF-Based EGFR Biochemical Assays

EGFR biochemical assays were carried out using a homogeneoustime-resolved fluorescence (HTRF) assay as described previously. Thereaction mixtures contained 1 μM biotin-Lck-peptide substrate, wild typeor mutant EGFR enzyme in reaction buffer (50 mM HEPES pH 7.1, 10 mMMgCl₂, 0.01% BSA, 1 mM TCEP and 0.1 mM Na₃VO₄) at a final volume of 10μL. Enzyme concentrations were adjusted to accommodate varying kinaseactivity and ATP concentrations (0.2-0.4 nM L858R/T790M; or 2-4 nML858R, or 2-4 nM T790M, or 40 nM WT). All reactions were carried out atroom temperature in white ProxiPlate™ 384-well Plus plates (PerkinElmer)and were quenched with 5 μL of 0.2 M EDTA at 60 min. Five μL per well ofthe detection reagent containing 2.5 ng PT66K (Cis-bio) and 0.05 μg SAXL(Prozyme) were added, and the plates were then incubated at roomtemperature for 1 hour and read with an EnVision plate reader. For IC₅₀determinations, compounds of the present application were diluted intoassay mixture (final DMSO 0.5%), and IC₅₀ values were determined by12-point inhibition curves (from 50 to 0.000282 μM) in duplicate underthe assay conditions as described above.

H1975, H3255 & HaCaT Target Modulation Assays Tissue Culture

Cells were maintained in 10% FBS/RPMI supplemented with 100 μg/mLPenicillin/Streptomycin (Hyclone #SH30236.01). The cells were harvestedwith 0.25% Trypsin/EDTA (Hyclone #SH30042.1), re-suspended in 5%FBS/RPMI Pen/Strep and plated at 7,500 cells per well in 50 μL of mediain a 384-well black plate with clear bottoms (Greiner #789068G). Thecells were allowed to incubate overnight in a 37° C., 5% CO₂ humidifiedtissue culture incubator. The 12-point serial diluted test compoundswere transferred to the plate containing cells by using a 50 nL Pin Headdevice (Perkin Elmer) and the cells were placed back in the incubatorfor 3 hours.

Phospho-EGFR (Y1173) Target Modulation Assay

HaCaT cells were stimulated with 10 ng/mL EGF (Peprotech # AF-100-15)for 5 minutes at room temperature. Constitutively activated EGFR mutantcell lines (H1975 and H3255) were not stimulated with EGF. The media wasreduced to 20 μL using a Bio-Tek ELx 405 Select™ plate washer. Cellswere lysed with 20 μL of 2× Lysis buffer containing protease andphosphatase inhibitors (2% Triton X-100, 40 mM Tris, pH 7.5, 2 mM EDTA,2 mM EGTA, 300 mM NaCl, 2× complete cocktail inhibitor (Roche #11 697498 001), 2× Phosphatase Inhibitor Cocktail Set II and Set III (Sigma#P5726 and #P0044)). The plates were shaken for 20 minutes. An aliquotof 25 μL from each well was transferred to prepared ELISA plates foranalysis.

For the experiment studying the effect of EGF pre-treatment on compound(e.g., compounds of the present application) target modulation, H1975cells were harvested and plated in 0.5% FBS/RPMI Pen/Strep. On thefollowing day, cells were pre-treated with 0.5% FBS/RPMI media with orwithout 10 ng EGF/mL for 5 minutes. Compound (i.e., compounds of thepresent application) was added and assay was carried out as describedabove.

Phospho-EGFR (Y1173) ELISA

Solid white 384-well high-binding ELISA plates (Greiner #781074) werecoated with μg/mL goat anti-EGFR capture antibody overnight in 50 mMcarbonate/bicarbonate pH 9.5 buffer. Plates were blocked with 1% BSA(Sigma #A7030) in PBS for 1 hour at room temperature, and washes werecarried out with a Bio-Tek ELx405 Select™ using 4 cycles of 100 μL TBS-T(20 mM Tris, 137 mM NaCl, 0.05% Tween-20) per well. A 25 μL aliquot oflysed cell was added to each well of the ELISA plate and incubatedovernight at 4° C. with gentle shaking. A 1:1,000 anti-phospho-EGFR in0.2% BSA/TBS-T was added and incubated for 2 hours at room temperature.After washing, 1:2,000 anti-rabbit-HRP in 0.2% BSA/TBS-T was added andincubated for 1 hour at room temperature. Chemiluminescent detection wascarried out with SuperSignal ELISA Pico substrate. Signal was read onEnVision plate reader using built-in UltraLUM setting.

Western Blotting

Cell lysates were equalized to protein content determined by CoomassiePlus™ Protein Assay Reagent (ThermoScientific #1856210) and loaded onto4-12% NuPAGE Bis-Tris gels with MOPS running buffer with LDS Samplebuffer (supplemented with DTT). Gel proteins were transferred to PVDFmembranes with an iBlot® Gel Transfer Device, 1× Casein-blockedmembranes were probed with primary antibodies overnight at 4° C. on anend-over-end rotisserie. Membranes were washed with TBS-T andHRP-conjugated secondary antibodies were added for 1 hour at roomtemperature. After washing, HRP was detected using Luminata™ ForteWestern HRP Substrate reagent and recorded with a Bio-Rad VersaDocimager.

Proliferation Assay

H1975, H3255 and HaCaT cell lines were plated in solid white 384-wellplates (Greiner) at 500 cells per well in 10% FBS RPMI P/S media. Usinga Pin Tool, 50 nL of serial diluted compounds of the present applicationwere transferred to the cells. After 3 days, cell viability was measuredby CellTiter-Glo (Promega) according to manufacturer's instructions.Luminescent readout was normalized to 0.1% DMSO-treated cells and emptywells. Data was analyzed by non-linear regression curve-fitting and EC₅₀values were reported.

Considering the allosteric mechanism of action the compounds of thepresent application, the extent to which ligand stimulation would affectpotency of inhibition of the mutant receptor was studied. To this end,inhibition of EGFR phosphorylation in H1975 cells in the presence andabsence of EGF using the quantitative ELISA-based assay was examined.

In the EGFR asymmetric dimer, the C-lobe of the “activator” subunitimpinges on the N-lobe of the “receiver” subunit, inducing an activeconformation in the receiver by reorienting the regulatory C-helix toits inward, catalytically functional position. In wild-type EGFR, onlythe receiver subunit is activated. Oncogenic mutations in the EGFRkinase domain induce an active conformation even in the absence ofligand stimulation, thus both subunits of a ligand-bound mutant receptorare expected to be catalytically active. In the receiver subunit but notthe activator, outward displacement of the C-helix is impeded by theasymmetric dimer interaction. Because the mutant receptor favors dimerformation and could promote dimerization even in the absence of ligand,this effect could explain the apparent disconnect in biochemical andcellular potencies of the allosteric inhibitor (Red Brewer et al., PNAS110, E3595-3604, doi:10.1073/pnas.1220050110 (2013); Shan et al., Cell149, 860 870, doi:10.1016/j.cell.2012.02.063 (2012)). To test thisnotion, an I941R point mutation in the C-lobe of the kinase, which isknown to block the asymmetric dimer interaction, was exploited. (Zhang2006; Cho et al., Cancer Res 73, 6770-6779,doi:10.1158/0008-5472.CAN-13-1145 (2013)). The activity of theL858R/T790M mutant is dimerization-independent, and as expected Ba/F3cells bearing the L858R/T790M/I941R triple mutant EGFR proliferated inthe absence of IL-3. The dimerization-defective mutant was dramaticallymore sensitive to the allosteric inhibitor.

One therapeutic antibody, cetuximab, targets the extracellular portionof the EGF receptor, blocking ligand binding and preventing dimerformation. The antibody is not effective clinically in EGFR-mutantNSCLC, and in cell-based studies cetuximab alone does not inhibitL858R/T790M or Del/T790M mutant EGFR, because their activity isdimerization independent.

Mouse Efficacy Studies

EGFR-TL (T790M/L858R) and EGFR-TD (exon 19 deletion-T790M) mice weregenerated as previously described. The EGFR-L858R:T790M:C797S (“TLCS”)mutant mouse cohort was established briefly as follows: The full-lengthHuTLCS cDNA was generated by site-directed mutagenesis using theQuickchange site directed mutagenesis kit (Agilent Technologies) andfurther verified by DNA sequencing. Sequence-verified targeting vectorswere co-electroporated with an FLPe recombinase plasmid into v6.5C57BL/6J (female)×129/sv (male) embryonic stem cells (Open Biosystems)as described elsewhere. Resulting hygromycin-resistant embryonic stemclones were evaluated for transgene integration via PCR. Then,transgene-positive embryonic stem clones were injected into C57BL/6blastocysts, and the resulting chimeras were mated with BALB/c WT miceto determine germline transmission of the TLCS transgene. Progeny of TL,TD and TLCS mice were genotyped by PCR of tail DNA.

The TL and TD mice were fed a doxycycline diet at 6 weeks of age toinduce EGFR TL or TD expression, respectively. The TLCS mice wereintranasally instilled with Ad-Cre (University of Iowa viral vectorcore) at 6 weeks of age to excise the loxP sites, activating EGFR TLCSexpression.

All care of experimental animals was in accordance with Harvard MedicalSchool/Dana-Farber Cancer Institute (DFCI) institutional animal care anduse committee (IACUC) guidelines. All mice were housed in apathogen-free environment at a DFCI animal facility and handled instrict accordance with Good Animal Practice as defined by the Office ofLaboratory Animal Welfare.

In Vivo Treatment and MRI Tumor Volume Quantification

The TL, TD and TLCS mice were monitored by MRI to quantify lung tumorburden before being assigned to various treatment study cohorts. All thetreatment mice had equal amount initial tumor burden. A compound of thepresent application was dissolved in 10% NMP (10%1-methyl-2-pyrrolidinone: 90% PEG-300), and was dosed at 60 mg/kg dailyby oral gavage. Cetuximab was administrated at 1 mg/mouse every threedays by intraperitoneal in injection. MRI evaluation was repeated every2 weeks during the treatment. The animals were imaged with a rapidacquisition with relaxation enhancement sequence (TR=2000 ms, TEeffect=25 ms) in the coronal and axial planes with a 1-mm slicethickness gating with respiratory rates. The detailed procedure for MRIscanning has been previously described (Li et al., 2007). The tumorburden volumes were quantified using 3-dimensional Slicer software.

Table 1 below shows the biological activity of representative compoundsof the application.

TABLE 1 Biochemical inhibitory activity (HTRF, IC₅₀) against recombinantEGFR T790M/L858R kinase, and antiproliferative activity (EC₅₀) againstT790M/L858R Ba/F3 cells in the absence and presence of 1 μg/mLcetuximab. Ba/F3 cellular activity (EC₅₀) HTRF (IC₅₀) T790M/L858R +Compound ID T790M/L858R T790M/L858R Cetuximab I-1 D D D I-2 D D D I-3 DD D I-4 D D D I-5 D D D I-6 D D D I-7 D D D I-8 D D D I-9 D D D I-10 A DA I-11 A D A I-18 D D D I-13 D D D I-14 A D A 0 < A < 250 nM; 250 nM ≤ B< 500 nM; 500 nM ≤ C < 750 nM; 750 nM ≤ D.

Example 19: Additional/Alternative Biochemical/Biological Studies CellViability Assays

H3255GR cells were treated with increasing concentrations of inhibitorsfor 72 hours and growth or the inhibition of growth was assessed by MTSassay according to previously established methods (Engelman et al.,2006; Ercan et al., 2015; Thou et al., 2009). All experimental pointswere set up in six technical replicates and all experiments wererepeated at least three times.

Western Blotting

To assess the effect of compounds on EGFR and its downstream pathways,NIH-3T3, H1975, H3255GR cells were treated for 4 hours before cells werelysed with NP40 lysis buffer, supplemented with protease and phosphataseinhibitors, followed by protein quantification. 20 μg of lysates wereused for Western Blotting analyses. For experiments that examine theeffect of an allosteric EGFR inhibitor in the presence of EGF, cellswere treated with 10 ng/ml of EGF for 15 minutes before they weretreated with drugs for 4 hours followed by lysis and proteinquantification as described above. All experiments were done at leastthree times.

Biotinylated Drug Pull Down Assay

For in vitro pull down assays, cells were treated with dose-escalatedWZ-4002, an ATP-competitive EGFR inhibitor for two hours before theywere subjected to lysis and protein quantification. 15-20 μg of proteinslysates were aliquoted and loaded at the same time as the pull downassay to ensure the presence of EGFR protein, phospho-EGFR activity.Tubulin expression was assessed to ensure even loading of gels. 500 μgof protein was incubated with either biotinylated-linker (control) orwith biotinylated allosteric EGFR inhibitor for two hours before 50%NeutrAvidin agarose beads (Thermo Fisher Scientific) slurry was addedfor an hour to precipitate the EGFR that was associated to thebiotinylated allosteric inhibitor. The beads were then washed threetimes with PBS containing 1% IGEPAL and an insulin syringe was used toremove extraneous buffer before the samples were suspended in 2×SDSsample preparation buffer for Western blotting analyses. All experimentswere performed at least three times.

ENU Mutagenesis

N-ethyl-N-nitrosourea (ENU) was purchased from Sigma Aldrich andmutagenesis studies were carried as previously described (Ercan et al.,2015). Briefly, 1×10⁶ cells/ml of L858R and L858R/T790M Ba/F3 cells weretreated with 50 μg/ml of ENU for 24 hours before the cells were washedthree times in RPMI media and expanded for 3 days. 1×10⁴ cells per wellwere plated in 96 wells and 5 plates were plated per condition. Thesecells were treated continuously with either DMSO, 1 μM gefitinib, 1 μMof an ATP-competitive EGFR inhibitor, 10 μM of an allosteric EGFRinhibitor alone or with gefitinib/allosteric EGFR inhibitor orATP-competitive EGFR inhibitor/allosteric EGFR inhibitor drugcombinations for 4 weeks with media and drug change once a week. Cellgrowth was monitored and number of resistant clones were counted andexpanded.

IncuCyte Studies

For cell confluency studies, H3255GR cells were treated with differentinhibitors and monitored by the automated microscopy using the IncuCyteLive-Cell Imaging system (Essen Bioscience). Confluency was measured byaveraging the percentage of area that the cells occupied from threeimages of a given well every two hours for 72 hours. For apoptosisstudies, cells were treated with inhibitors incubated in mediacontaining the CellEvent™ Caspase 3/7 Green ReadyProbes® reagent (ThermoFisher Scientific) and monitored for change in green fluorescenceactivity using the aforementioned imaging system. The average number ofobjects that were stained with green from three images per well wascounted as positive for Caspase 3/7, indicating apoptosis, and recordedevery two hours for 72 hours. All experimental conditions were set up inat least six replicates and all experiments were performed at leastthree times.

In Vivo Studies

All breeding, mouse husbandry, and in vivo experiments were performedwith the approval of the Dana-Farber Cancer Institute (Boston, Mass.)Animal Care and Use Committee.

For the H1975 xenograft study, Nu/Nu mice were purchased from CharlesRiver Laboratories International Inc. H1975 cells were detected aspathogen free at Charles River Laboratories International Inc. and wereresuspended in serum-free medium mixed with an equal amount of Matrigel(BD Biosciences). Mice were injected at 2 locations per mouse in theflanks with 2 million cells per shot. The mice were randomly grouped,and treatment started when tumor size reached 100 to 200 mm³. Eachcohort included at least 5 mice. Tumor sizes were monitored weekly, andvolumes were calculated using the following formula:(mm³)=length×width×width×0.5.

To assess EGFR activity in the mice after the study was performed,tumors were taken 3 hours after the last dose for pharmacodynamic (PD)studies. Tumors were flash frozen in liquid nitrogen to preserve tissueintegrity and homogenized in RIPA buffer supplemented with protease andphosphatase inhibitors. The protein was quantified and 20 μg of lysateswere used for Western Blotting analyses.

In the H1975 xenograft study, an allosteric EGFR inhibitor was dissolvedin 5% NMP (5% 1-methyl-2-pyrrolidinone: 95% PEG-300). An allosteric EGFRinhibitor was dosed at 100 mg/kg once daily orally. An ATP-competitiveEGFR inhibitor was dissolved in 0.5% HMPC (0.5% Hydroxypropylmethylcellulose: 99.5% 0.05N hydrogen chloride). Mice received 25 mg/kgATP-competitive EGFR inhibitor once daily orally.

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain, usingno more than routine experimentation, numerous equivalents to thespecific embodiments described specifically herein. Such equivalents amintended to be encompassed in the scope of the following claims.

1. A compound of Formula I:

or a pharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof, wherein: A₁ is phenyl or 5- or 6-membered heteroaryl comprising1-3 heteroatoms selected from N, O, and S, wherein the phenyl orheteroaryl is optionally substituted with one or more R₀; each R₀ isindependently C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆ hydroxyalkyl, C₁-C₆alkoxy, C₁-C₆ haloalkoxy, OH, halogen, CN, phenyl, C₃-C₆ cycloalkyl, 5-or 6-membered heteroaryl comprising 1-3 heteroatoms selected from N, O,and S, or 5- or 6-membered heterocyclyl comprising 1-3 heteroatomsselected from N, O, and S, wherein the phenyl, cycloalkyl, heteroaryl,or heterocyclyl is optionally substituted with one or more substituentsindependently selected from C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆hydroxyalkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkoxy, OH, and halogen, or twoR₀, taken together with the adjacent atoms to which they are attached,form phenyl, C₅-C₆ cycloalkyl, 5- or 6-membered heteroaryl comprising1-3 heteroatoms selected from N, O, and S, or 5- or 6-memberedheterocyclyl comprising 1-3 heteroatoms selected from N, O, and S,wherein the phenyl, cycloalkyl, heteroaryl, or heterocyclyl isoptionally substituted with one or more substituents independentlyselected from C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆ hydroxyalkyl, C₁-C₆alkoxy, C₁-C₆ haloalkoxy, OH, and halogen; R₁ is H, C₁-C₆ alkyl, C₁-C₆haloalkyl, C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkoxy, or(CH₂)_(m)-A₂; A₂ is phenyl, C₃-C₆ cycloalkyl, 5- or 6-memberedheteroaryl comprising 1-3 heteroatoms selected from N, O, and S, or 5-or 6-membered heterocyclyl comprising 1-3 heteroatoms selected from N,O, and S, wherein the phenyl, cycloalkyl, heteroaryl, or heterocyclyl isoptionally substituted with one or more substituents independentlyselected from C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆ hydroxyalkyl, C₁-C₆alkoxy, C₁-C₆ haloalkoxy, OH, and halogen; m is 0, 1, 2, or 3; R₂ is Hor C₁-C₆ alkyl; R₃ is H, halogen, C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆hydroxyalkyl, C₁-C₆ alkoxy, or C₁-C₆ haloalkoxy; or R₂ and R₃, takentogether with the intervening atoms, form a 5- or 6-memberedheterocyclyl ring comprising 0-2 additional heteroatoms selected from N,O, and S, or a 5- or 6-membered heteroaryl ring comprising 0-2additional heteroatoms selected from N, O, and S, wherein theheterocyclyl or heteroaryl is optionally substituted with one or moresubstituents independently selected from oxo, halogen, C₁-C₆ alkyl,C₁-C₆ haloalkyl, C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy, and C₁-C₆ haloalkoxy;L₁ is a bond, a straight or branched bivalent C₁-C₆ alkylene chain,—C(O)—, —C(O)(CH₂)_(n)—, —C(O)NH—, —C(O)NH(CH₂)_(n)—, —C(O)N(C₁-C₆alkyl)-, —C(O)N(C₁-C₆ alkyl)(CH₂)_(n)—, —C(O)O—, —C(O)O(CH₂)_(n)—,—C(S)—, —C(S)(CH₂)_(n)—, —C(S)NH—, —C(S)N(C₁-C₆ alkyl)-,—C(S)NH(CH₂)_(n)—, —C(S)N(C₁-C₆ alkyl)(CH₂)_(n)—, —(CH₂)_(p)NH—,—(CH₂)_(p)N(C₁-C₆ alkyl)-, or —(CH₂)_(p)O—; n is 1 or 2; p is 1, 2, or3; X₁, X₂, X₃, and X₄ are each independently N or CR₄, provided that atleast two of X₁, X₂, X₃, and X₄ are CR₄; each R₄ is independently H,NR₅R₆, NR₇C(O)R₈, C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆ hydroxyalkyl,C₁-C₆ alkoxy, C₁-C₆ haloalkoxy, OH, halogen, CN, phenyl, C₃-C₆cycloalkyl, 5- or 6-membered heteroaryl comprising 1-3 heteroatomsselected from N, O, and S, or 5- or 6-membered heterocyclyl comprising1-3 heteroatoms selected from N, O, and S, wherein the phenyl,cycloalkyl, heteroaryl, or heterocyclyl is optionally substituted withone or more substituents independently selected from C₁-C₆ alkyl, C₁-C₆haloalkyl, C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkoxy, OH, andhalogen; each R₅ and each R₆ are independently H or C₁-C₄ alkyl; each R₇is independently H or C₁-C₄ alkyl; each R₈ is independently C₁-C₄ alkyl;X₅, X₆, X₇, and X₈ are each independently N, CR₉, or CR₁₀, provided thatat least one of X₅, X₆, X₇, and X₈ is CR₉ and at least one of X₅, X₆,X₇, and X₈ is CR₁₀; each R₉ is independently H, halogen, OH, CN, C₁-C₆alkyl, C₁-C₆ haloalkyl, C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy, or C₁-C₆haloalkoxy; each R₁₀ is independently phenyl, C₃-C₆ cycloalkyl, 5- or6-membered heteroaryl comprising 1-4 heteroatoms selected from N, O, andS, or 5- or 6-membered heterocyclyl comprising 1-3 heteroatoms selectedfrom N, O, and S, wherein the phenyl, cycloalkyl, heteroaryl, orheterocyclyl is optionally substituted with one or more R; each R₁₁ isindependently C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆ hydroxyalkyl, C₁-C₆alkoxy, C₁-C₆ haloalkoxy, OH, halogen, CN, phenyl, C₃-C₆ cycloalkyl, 5-or 6-membered heteroaryl comprising 1-3 heteroatoms selected from N, O,and S, or 5- or 6-membered heterocyclyl comprising 1-3 heteroatomsselected from N, O, and S, wherein the phenyl, cycloalkyl, heteroaryl,or heterocyclyl is optionally substituted with one or more substituentsindependently selected from C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆hydroxyalkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkoxy, OH, and halogen, or twoR₁₁, taken together with the adjacent atoms to which they are attached,form phenyl, C₅-C₆ cycloalkyl, 5- or 6-membered heteroaryl comprising1-3 heteroatoms selected from N, O, and S, or 5- or 6-memberedheterocyclyl comprising 1-3 heteroatoms selected from N, O, and S,wherein the phenyl, cycloalkyl, heteroaryl, or heterocyclyl isoptionally substituted with one or more substituents independentlyselected from C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆ hydroxyalkyl, C₁-C₆alkoxy, C₁-C₆ haloalkoxy, OH, and halogen.
 2. The compound of claim 1,wherein A₁ is phenyl optionally substituted with one or more R₀. 3-4.(canceled)
 5. The compound of claim 1, wherein L₁ is —C(O)—,—C(O)(CH₂)_(n)—, —C(O)NH—, —C(O)NH(CH₂)_(n)—, —C(O)N(C₁-C₆ alkyl)-, or—C(O)N(C₁-C₆ alkyl)(CH₂)_(n)—.
 6. (canceled)
 7. The compound of claim 1,wherein X₁, X₂, X₃, and X₄ are each CR₄. 8-12. (canceled)
 13. Thecompound of claim 1, wherein at least one R₄ is halogen.
 14. Thecompound of claim 1, wherein one of X₅, X₆, X₇, and X₈ is CR₁₀, andthree of X₅, X₆, X₇, and X₈ are each independently CR₉. 15-16.(canceled)
 17. The compound of claim 1, wherein one of X₅, X₆, X₇, andX₈ is N, one of X₅, X₆, X₇, and X₈ is CR₁₀, and two of X₅, X₆, X₇, andX₈ are each independently CR₉. 18-19. (canceled)
 20. The compound ofclaim 1, wherein R₂ is H.
 21. (canceled)
 22. The compound of claim 1,wherein R₃ is H.
 23. (canceled)
 24. The compound of claim 1, wherein R₃is C₁-C₆ alkoxy. 25-26. (canceled)
 27. The compound of claim 1, whereinR₂ and R₃, taken together with the intervening atoms, form a 5-memberedheterocyclyl ring comprising 0-2 additional heteroatoms selected from N,O, and S, wherein the heterocyclyl is substituted with oxo. 28.(canceled)
 29. The compound of claim 1, wherein each R₉ is H.
 30. Thecompound of claim 1, wherein at least one R₉ is halogen, OH, CN, C₁-C₆alkyl, C₁-C₆ haloalkyl, C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy, or C₁-C₆haloalkoxy.
 31. The compound of claim 1, wherein at least one R₁₀ isphenyl optionally substituted with one or more R₁₁.
 32. (canceled) 33.The compound of claim 1, wherein at least one R₁₀ is 5- or 6-memberedheteroaryl comprising 1-4 heteroatoms selected from N, O, and Soptionally substituted with one or more R. 34-37. (canceled)
 38. Thecompound of claim 1, wherein at least one R₁₀ is 5- or 6-memberedheterocyclyl comprising 1-3 heteroatoms selected from N, O, and Soptionally substituted with one or more R₁₁. 39-44. (canceled)
 45. Thecompound of claim 1, wherein R₁ is H.
 46. (canceled)
 47. The compound ofclaim 1, of Formula IIa′ or IIg′:

or a pharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof, wherein: b is 0, 1, or 2; and c is 1, 2, or
 3. 48-53.(canceled)
 54. The compound of claim 1, of Formula IVa′

or a pharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof. 55-70. (canceled)
 71. The compound of claim 1, selected fromthe group consisting of:

or a pharmaceutically acceptable salt, hydrate, solvate, or stereoisomerthereof.
 72. A pharmaceutical composition comprising a compound of claim1, or a pharmaceutically acceptable salt, hydrate, solvate, orsteroisomer thereof, and a pharmaceutically acceptable carrier,optionally further comprising a second agent that prevents EGFR dimerformation, and a pharmaceutically acceptable carrier.
 73. A kitcomprising a compound of claim 1, or a pharmaceutically acceptable salt,hydrate, solvate, or stereoisomer thereof, optionally further comprisinga second agent that prevents EGFR dimer formation, and apharmaceutically acceptable carrier.
 74. (canceled)
 75. A method oftreating or preventing a disease, a disease resistant to an EGFRtargeted therapy, cancer wherein the cell of the cancer comprises anactivated EGFR or an activated ERBB2, or cancer in a subject wherein thesubject is identified as being in need of EGFR inhibition or ERBB2inhibition for the treatment or prevention of cancer, comprisingadministering to a subject in need thereof an effective amount of acompound of claim 1, or a pharmaceutically acceptable salt, hydrate,solvate, or stereoisomer thereof.
 76. The method of claim 74, furthercomprising administering a second agent that prevents EGFR dimerformation, and a pharmaceutically acceptable carrier. 77-84. (canceled)